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IVD medical equipment
Fluorescent Quantitative
Detection system
LineGene 9600 Series
User’s Manual
Attention Users are recommended to carefully read the contents of this manual before operating the
Bioer Fluorescent Quantitative PCR Detection System.
To carefully observe all special Warnings and Cautions outlined in this manual.
This manual should be maintained properly in good condition for reference.
Caution:
Copyright reserved. The Bioer Co. reserves the right to modify this manual at any time without notice.
The manual contains copyright protected and patented material. Without prior written consent from Hangzhou Bioer Technology Co., Ltd., any part of the manual shall not be duplicated, reproduced or translated into any other language.
Thank you for your purchase of this product.
Before initial use of this instrument, please carefully read this manual
!
File No.: BYQ60840000000ESM
File Version: January August AugustMarch May 2012 0 Version 1 ..186
0
LineGene9600 Important Notes
Important Notes
1. Usual practice
Announce:
Very important information is contained in any item and should be carefully read. Failure to operate instrument according to the instruction would result in damage or abnormal function of instrument.
Warning!
The warning message requires extremely careful operation of a certain step.
Failure to observe the instruction would result in serious personal injury.
2. Safety
During operation, maintenance and repair of this instrument, the following basic safety notes shall be observed. In case of failure to follow these measures or the warnings or notes indicated herein, the basic protection provided by the instrument, its safety criteria of design and manufacture, and its predicted use range would be impaired.
Hangzhou Bioer Technology Co., Ltd. shall be irresponsible for the consequence resulted from user’s failure to observe the following requirements.
Note:
The instrument, complying with the Standard GB4793.1
/IEC61010-1 , is a general instrument of classⅠ, the protection degree is IP20. It is intended for indoor use.
Note:
The instrument, complying with the Standard YY0648 /IEC61010-2-101 , is used for
IVD medical equipment.
a) Instrument earth
In order to avoid electrical shock accident, the input power wire of instrument shall be reliably earthed. This instrument uses 10A 3-core earth plug and this plug is provided with a third (earth) leg, is merely used for earth type power socket and is a safety unit. If the plug is impossible to insert into the socket, the socket shall be correctly fixed by electrician; otherwise, the earth plug would lose safe protection function.
b) Keeping apart from the live circuit
The operator shall not arbitrarily remove instrument protector, replace elements or make inside adjustment, and the task, if necessary, shall be made by qualified professional maintenance personnel. Replacement of elements under energizing state shall be prohibited.
c) Use of power supply
Before turning on AC power, make the power voltage consistent with the instrument-required one (100-240V~, 50/60Hz) and make the rated load power socket not below instrument’s maximum load 600W.
d) Power wire
I
LineGene9600 Important Notes
Generally, this instrument uses accompanied power wire. The damaged power wire shall be replaced by one of the same type and same specification. It can’t be repaired. The power wire shall neither be compressed nor be laid in gathered personnel location.
e) Insertion and withdrawal of power wire
At insertion and withdrawal of power wire, the operation portion of the plug shall be correctly held with hand. At insertion, the plug shall be completely and tightly inserted into the socket and at withdrawal heavily pulling the wire shall not be allowed.
f) Placement of instrument
This instrument should not be laid in the place where it is different to cut off the power supply.
This instrument shall be fixed in a low RH and low dust place apart from water source (e.g.
water pool and water pipeline) and the room shall be well ventilated, and free of corrosive gas or and interference of strong magnetic field. The instrument shall not be placed in a wet or dusty location and its working table shall be level and steady.
The openings on this instrument are for ventilation circulation and in order to avoid over-head of the instrument body they shall not be blocked or covered. When a single set or several sets of instrument are used, the interval between its surrounding ventilation opening and its nearest object shall not be below 30cm or 50cm.
Excessive environmental temperature would impair the test performance or result in fault of instrument. This instrument shall not be used in location subjected to direct sunshine or strong radiation of light source, otherwise, its fluorescence detection would be impaired. The instrument shall be kept away from hot gas, strove and all other heat source.
At shutdown, the power shall be cut off. If the instrument is to be set aside for a long time, the power shall be cut off, the power plug withdrawn and the instrument covered with soft cloth or plastic film to avoid entrance of dust and foreign matters.
g) Notes during operation
During test, cares shall be taken to prevent liquid from dropping onto the instrument. The castoff used in test , such as consumables, reagent, and so on, should be treated as require, and should not be thrown away or poured.
During test, if there is hazardous substances, user must be trained before using.
Hazardous substances, which has been used, should be coped with and saved according to derection for use.
User, who operates the instrument, must be trained and has relevant quantification.
Caution:
In any of the following cases, immediately cut off the power supply, withdraw the power plug from the power socket, contact the supplier or ask qualified repair personnel for treatment:
Liquid drops into inside of the instrument.
The instrument is rained or watered.
The instrument works abnormally, especially generates abnormal sound or odor.
II
LineGene9600 Important Notes
The instrument is dropped or its casing is damaged.
The function of instrument obviously changes.
Caution:
When you deal with potential contagious matter such as flesh sample or reagent, which is likely to touch skin, protecting glove or other protecting measures are need to be used.
h) Transportation again
If transporting the instrument again, the instrument and its detection wells need be cleared wholely before transportation, and be disinfected by UV light. After clearing, the fixing pin shall be tightened to transportation well when the instrument stops running and power off so that motion componet of the instrumnt interior can be fixed and motion componet is avoided to be demaged during transportation. Introduce of the location from transportation well is seen in chapter of fixing pin backout.
i) Warning Signmark
Warning identification
DANGER!
SCALDING!
Place pasted this mark in instrument, is danger, if the instrument is used irrelevancy.
Place pasted this mark in instrument, causes high temperature and is scalding during use.
BIOHAZARD
PROTECTIVE
EARTH
Place pasted this mark in instrument, caused biohazard during use.
Protective earth is near to the place pasted this mark in instrument
Warning mark
Warning!
The instrument is adhered with mark “HOT SURFACE!” During operation of instrument or a period immediately after completion of running of a program, any portion of body shall not be contacted with metallic portion (module) nearby the said mark; otherwise, skin would be burnt.
Warning!
Operator may touch matter which is baneful to organism or is contagious during using instrument. Operate should realize the harmfulness, and operate in the condition which must accord with rules of PCR lab. And operator must be
III
LineGene9600 trained or be provide with qualification.
Important Notes j) Sign of external packing
Up
Breakables
Fear rain
The limit of stacking layer
Temperature limit
Correct position of transport package is vertical upwards.
Be treated with care when transportation, there is breakables in transport package,
Transport package is not rained.
Maximum stacking layer of the same package is 2.
Temperature limit that transport package should keep is form -20C to 55C.
3. EMC Consideration
Note:
This is a class A equipment, only suitable for use in establishments other than domestic, and those directly connected to a low voltage power supply network which supplies buildings used for domestic purpose.
4. Maintenance of instrument
The stained surface of this instrument, if any, shall be cleaned with soft cloth soaked with cleaning paste.
Heat conductive oil medium shall not be used in module holes of this instrument.
Module should be pushed in in time to prevent dust during normal conservation or after use.
Warning!
At cleaning of instrument, its power supply shall be cut off.
The instrument surface shall not be cleaned with corrosive cleaning agent.
Instrument module includes precise optics, dust, foreign matter and residue should be avoided.
5. After-sales services
The warranty content and scope are shown in the warranty sheet.
IV
LineGene9600 Important Notes
Note:
After unpacking, immediately check the goods against the packing list. In case of damaged or missed goods, please immediately contact the supplier.
After qualification of acceptance, properly fill the product acceptance sheet and send (or fax) the copied sheet to the supplier for filing and maintenance.
Before use of the product, the user shall feedback the information registration table to Hangzhou Bioer Technology Co., Ltd. and obtain correct operation password.
After unpacking, the packing box and packing materials shall be well kept for future repair.
The instrument should be disinfected before sent to the repair department.
Fixing pin must be fixed correctly as manual before transporting instrument.
Maintainer must disinfect the instrument immediately after unpacking when instrument is sent to repair department.
For the damage of instrument occurring during transporting to the repair department due to improper package or incorrect place of fixing pin,
Hangzhou Bioer Technology Co., Ltd. shall bear no liability.
V
LineGene9600 Contents
Contents
CHAPTER 1 GENERAL DESCRIPTION...............................................................................................1
1. A
PPLICATIONS
........................................................................................................................................ 1
2. F
EATURES
...............................................................................................................................................1
3. N
OTES TO MODEL AND DESCRIPTION
..................................................................................................... 2
4. P
ERFORMANCE PARAMETERS
.................................................................................................................2
5. G
ENERAL DESCRIPTION OF FUNCTIONS OF THE NECESSARY SOFTWARE
................................................ 3
CHAPTER 2 PREPARATIONS................................................................................................................ 4
1. T
RANSPORT AND STORAGE CONDITION OF INSTRUMENT
.......................................................................4
2. N
ORMAL WORKING CONDITION
..............................................................................................................4
3. P
REPARATIONS BEFORE START OF INSTRUMENT
....................................................................................4
3.1 Removal of fixing pin..................................................................................................................... 4
3.2 Connection of power wire and communication wire......................................................................5
4. S
YSTEM
I
NSTALLATION AND
U
NLOADING
............................................................................................. 5
4.1 System Installation.......................................................................................................................... 5
4.2 System Unloading........................................................................................................................... 5
CHAPTER 3 START...................................................................................................................................6
1. C
HECKS BEFORE START
..........................................................................................................................6
2. S
TART
.....................................................................................................................................................6
3. S
TARTING SOFTWARE INTERFACE
........................................................................................................... 6
CHAPTER 4 ABSOLUTE QUANTIFICATION..................................................................................... 8
1. D
ESIGN
E
XPERIMENT
.............................................................................................................................8
1.1 Create New Absolute Quantitative Experiment...........................................................................9
1.2
Detector Setting.........................................................................................................................9
1.3 Sample Information Setting....................................................................................................... 10
1.4 Reaction Plate Setting................................................................................................................ 11
1.5 Program Setting..........................................................................................................................13
2. P
REPARE FOR
R
EACTION
......................................................................................................................15
3. R
UN THE
E
XPERIMENT
.........................................................................................................................16
3.1 Preparation for reagent sample......................................................................................................17
3.2 Start Run........................................................................................................................................17
3.3 Monitor Operation.........................................................................................................................18
3.4 Program Setting.............................................................................................................................19
4. E
XPERIMENT
A
NALYSIS
....................................................................................................................... 21
4.1 Check Results................................................................................................................................ 22
4.2 Adjust Parameter Reanalysis.........................................................................................................27
5. E
XPERIMENT
R
EPORT
.......................................................................................................................... 29
5.1 Report Template Designing...........................................................................................................30
5.2 Print Setting...................................................................................................................................30
i
LineGene9600 Contents
5.3 Report Printing.............................................................................................................................. 31
6. D
ATA
E
XPORT
...................................................................................................................................... 32
6.1 Export to Database........................................................................................................................ 33
6.2 Experiment Filing..........................................................................................................................33
6.3 Export Experiment Data to EXCEL..............................................................................................33
CHAPTER 5 TOOL USAGE.................................................................................................................... 34
1. G
AIN
S
ETTING
......................................................................................................................................34
2. T
ARGET
L
IBRARY
.................................................................................................................................34
3. C
USTOMIZED
D
YES
...............................................................................................................................34
4. C
USTOMIZE
C
OLUMNS
......................................................................................................................... 35
5. C
OLUMN
S
ELECTION
............................................................................................................................36
6. S
AMPLE
C
OLUMN
L
IBRARY
................................................................................................................. 36
7. I
NSTRUMENT
C
ALIBRATION
P
ARAMETERS
...........................................................................................37
8. M
EASURE
C
ROSSTALK
C
ALIBRATION
P
ARAMETERS
........................................................................... 37
9. C
ROSSTALK
G
AIN
P
ARAMETER
M
EASUREMENT
................................................................................. 38
10. S
YSTEM
M
AINTENANCE
..................................................................................................................... 39
11. F
IRMWARE
U
PGRADING
..................................................................................................................... 39
12. C
ONVERT
O
LD
F
ILE INTO
N
EW
F
ILE
..................................................................................................40
CHAPTER 6 OTHER FUNCTIONS.......................................................................................................41
1. I
NSTRUMENT
O
PERATION
.....................................................................................................................41
1.1 Connect Instrument....................................................................................................................... 41
1.2 Disconnect..................................................................................................................................... 41
1.3 Instrument Information................................................................................................................. 41
2. D
ATA
Q
UERY
........................................................................................................................................41
3. O
PEN AND
S
AVE
F
ILE
...........................................................................................................................42
4. S
YSTEM
H
ELP
.......................................................................................................................................42
5. A
BOUT
..................................................................................................................................................42
CHAPTER 7 MAINTENANCE............................................................................................................... 43
1. R
EGULAR CLEANING
............................................................................................................................43
2. A
NALYSIS AND ELIMINATION OF TROUBLES
........................................................................................ 43 ii
LineGene9600 General Description
Chapter 1 General description
This chapter mainly describes the applications, features, specification, model, performance parameters and software function of 96-well Fluorescent Quantitative Detection System.
1. Applications
96-well fluorescent quantitative PCR detection system adopts amplification of fluorescent real-time detection mode analysis PCR module and is suitable for polymerase chain reaction fluorescent quantitative detection in research fields of human genome engineering, forensic, cancer, tissue, population biology, paleontology, zoology and botany and in clinical diagnosis of virus, cancer and genie diseases.
PCR detection system belongs to IVD medical equipment, which is to use polymerase chain reaction means to quantitative analysis of different genes in clinic lab.
2. Features
Novel and human-orientated running and operation interface make operation much smooth.
The adopted fluorescent real-time detection mode realizes simultaneous amplification and detection in a same tube without need of post treatment.
Advanced thermoelectric refrigeration technology ensures fast and steady heating and refrigerating of ultra-fast heat cycles system.
Multi-point temperature control ensures much even temperature of 96 sample holes.
It can control the temperature gradient of 4 areas through setting.
Stable and correct 1~36°C gradient function makes PCR condition optimal.
The constant temperature function of SOAK realizes low-temperature storage of PCR reagent.
It uses maintenance-free LED excitation light source of long service life.
The advanced optic fiber transmission technology makes photo-electric detection system much sensitive and reliable.
Precise optical path system and ultra-sensitive PMT system render best correctness and sensitivity of fluorescent detection.
It can make real-time dynamic monitor of the entire PCR amplification process.
It has high linear range up to 10 orders of start DNA copies without gradient dilution.
PCR reaction tube is unnecessary to open, which protects products during and after PCR from contamination and ensures correct results.
The multi-color fluorescence monitor may acquire much information from a single reaction.
The hot-lid technology realizes oil-free operation of PCR.
Automatic hot-lid technology needs no manual opening/closing and ensures constant pressure of hot-lid used in different-high test tube.
It has full-Chinese interface and the functions of flexible setting of program and overall analysis and report and can store all parameters.
It can print out several or a single sample report.
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LineGene9600 General Description
Automatic, correct and timely remote networked services provide 96-well quantitative detection system the latest technical support.
The advanced module bottom fluorescent detection technology is protected by invention patent.
3. Notes to model and description
Model:
FQD – 96 A
Description:
LineGene 96 ××
Design series number
Specification code, 96 denoting maximum sample quantity 96
Abbreviation for Fluorescent quantitative polymerase chain reaction (PCR) detection system
Code of ray filter pack
20 (with ray filter of F1,F2)
40 (with ray filter of F1,F2,F3,F4)
60 (with ray filter of F1,F2,F3,F4,F5,F6)
80 (with ray filter of F1,F2,F3,F4,F5,F6,F7,F8)
Specification code, 96 denoting maximum sample quantity 96
English name of description of Bioer Fluorescent quantitative polymerase chain reaction (PCR) analysis system series.
4. Performance parameters
Specification/ model
Sample capacity
Detection channel
FQD-96A
96×0.2ml (suitable for single tube, 8-row tubes and 96-well full-apron plate)
F1 F2 F3 F4 F5 F6 F7
F8
-
Applicable dye
FAM,
SYBR
Green I
VIC,
HEX,
TET,
JOE
Cy3,
5-TAMRA,
NED
ROX,
TEXAS-R
ED
Cy5
Light
Cycler Red
Quasar 705
-
Temperature range of block working
4~105°C (Minimum division:0.1°C)
Heating/cooling
-2-
4.0°C/s (max)
LineGene9600 General Description rate
Temperature fluctuation
Temperature accuracy
Temperature uniformity
Temperature range of hot-lid working
Repeatability of fluorescent intensity detection
Running mode
Operation system
Power supply
≤
≤
0.1°C (full-range), (55°C typical value ≤0.1°C)
0.2°C (full-range), (55°C typical value ≤0.1°C)
≤0.4°C (full-range), (55°C typical value ≤0.3°C)
30~110°C (adjustable, default 105°C)
5%
Continuous running
Windows2000/XP/Windows Vista/Windows7
100-240V~ 50/60Hz 600W
Dimensions 430mm×395mm×352 mm
Weight 28kg
Caution: F7 and F8 are the preserved dummy channels and may be customized as per actual requirements of customers.
5. General description of functions of the necessary software
a) Parameters setting-up function (including temperature, time, cycles, heating/cooling rate, selection of detection channel and yield of photo-electric amplification tube) b) Note function of text contents c) Sample material record function (sample No., sample name and sample data) d) Document running display function (PCR heat cycle data display, fluorescence detection data display and real-time display of each data during running of instrument) e) Detection data analysis function (The analysis function may be independently used under no connection of instrument) f) Analysis result output function (It may output the analysis result to other type document, e.g.:
EXCEL, TXT document, it can inquiry and print out analysis result, modify printing format and select printing items.) g) Document storage function (setting up data, running data and analysis result) h) Trouble protection and alarm function
Caution:
The above-mentioned software function is merely for reference. The software function would be modified without notice.
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LineGene9600 Preparations
Chapter 2 Preparations
This chapter mainly describes use, transport and storage condition, structural composition, removal of fixing pin, installation/unloading of software and preparations before start of
LineGene 9600 series fluorescent quantitative detection system.
1. Transport and storage condition of instrument
Environmental temperature: - 20°C~55°C
Relative Humidity: ≤80%
2. Normal working condition
Environmental temperature: 10°C~30°C
Environmental RH: ≤70%
Power supply: 100-240V~ 50/60Hz 600W
Caution:
Before use the instrument, please make sure the working condition meets the above requirements. The power socket shall be 3-holesocket and reliably earthed.
3. Preparations before start of instrument
3.1 Removal of fixing pin
In order to prevent the inner moving parts from shift and collision during transport, before leaving the factory they are fixed with fixing pin. After the instrument is placed on the desired location, the fixing pin shall be taken out through counterclockwise rotation, inserted to unlocker hole at its back, and fully tightened to release protection of the inside protective switch. Only after correct installation of the fixing pin, may the instrument be energized; otherwise, the instrument’s temperature control system is able to run but X-Y axis scanning system doesn’t run.
Fixing pin
Can be removed through rotation
Please keep the fixing pin tightened. Before moving instrument, turn on the power, pending
-4-
LineGene9600 Preparations completion of self-test action ,then shut down instrument normally and take out the fixing pin, insert it into right hole of instrument and tighten it.
3.2 Connection of power wire and communication wire
Connection of power wire: The instrument-accompanied power wire should be used. At connection the instrument power switch shall be at “OFF” state and after connection the power wire shall be checked for tight contact with the instrument socket; otherwise it shall be replaced.
Connection of communication wire: The accompanied communication wire and communication conversion box shall be used. One end of the communication conversion box shall be connected with DB15 communication interface at instrument back and another end connected with computer USB interface or RS232C interface or Bluetooth interface . After connection, screws shall be surely tightened.
Caution:
The accompanied power wire is reliable, but after several times of insertion and withdrawal the connection would likely become loose. In this case, the power wire shall be replaced with the one of same type and same specification.
The communication conversion box is built with special circuits and shall not be arbitrarily opened without permit.
4. System Installation
and Unloading of necessary software
4.1 System Installation
System Environment
Operating system: Windows XP/Windows VISTA/Windows7
Runtime environment: Net Framework 4.0
Other software: PDF reader
Minimum configuration:
Processor: Intel Core i3
Memory: 2GB
Hard Disc: 10GB
System installation
Double click LineGene9600 installation file (
LineGene9600DiagnosisSetup.exe
) display the installation interface (select installation language) set up installation pathinstall
Operating system
Double click the LineGene9600 shortcut on the desktop
Click the start menuProgram LineGene9600
4.2 System Unloading
Control PanelAdd/Delete Program LineGene9600Unload
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LineGene9600 Preparations
Selection of computer system
Minimum configuration:
Operation system: Windows 2000/XP/Windows Vista/Windows7 ( English professional or ultimate)
Application software: Microsoft Excel2000/2002/2003,/2007
Microsoft Access2000/2002/2003/2007
EMS memory: 512M
Hard disk: 10G
CPU: Pentium® 4
System setting up: virtual internal mummery above 1000MB
Installation of LineGene 9600 software
After computer starting Windows2000/XP/Windows Vista/Windows7system, insert the installation compact disk marked “LineGene 9600” into optical drive, open compact disk, enter
LineGene 9600 folder, double click setup.exe under this directory and enter language selection installation interface of LineGene 9600 fluorescent quantitative detection system software:
After selection of installation language click “OK ” to enter the next interface:
Click “Next >” and enter the next interface:
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LineGene9600 Preparations
Please input corresponding column: user name, organization and series number.
Caution: The installation series number of software is shown on the label on front of installation compact disk of LineGene 9600 Fluorescent Quantitative Detection System.
Click “Next >”and enter the following interface:
The screen displays installation target position (installation directory). The default installation directory is C:\Program Files\bioer\ LineGene 9600\. For confirmation of the said installation directory, click “Next >” and for modification or new establishment of installation directory, click
“Change…” and enter the following installation directory selection interface:
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LineGene9600 Preparations
On selection interface of the installation directory, from column “Search range ” select the desired installation route or from the column “Folder name” directly input or modify the installation target position, then click “OK”, successfully establish route target position and enter
“Custom Setup” interface. Select the program which you want to install.
Select the program which you want to install, click “Next >” enter automatic installation. After installation, LineGene 9600 series fluorescent quantitative detection system short-cut icon
(Which you selected.) will be automatically formed on desktop:
Only channels F1,F2 are allowed with LineGene9620.
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LineGene9600 Preparations
Only channels F1,F2,F3,F4 are allowed with LineGene9640.
Only channels F1,F2,F3,F4,F5,F6 are allowed with LineGene9660
All channels are allowed with LineGene9680
Selection of software installation depends on detection channels of instrument. When installation, you should select assorted software according to description of instrument.
After instrument is connected with computer successfully, there are description and number in the status bar, in which usable detection channels are displayed.
Caution1:All files of modification parameter are saved in folder
“AttachedFiles\LineGene9600_******” of root directory of disk. When user installing software, all files of modification parameter are copied automatically in installation directory.
Caution2
:
After installing the software on the computer without connected to instrument, before analysising the data,please copying default.ser file from the computer connected with instrument to the computer without connected with the instrument
(
the default .ser file was automatically generated by the computers connected with the instrument can’t be analysed correctly.
Unloading of LineGene 9600 software
In [Start]/ [Setup]/ [Control panel] menu click the “Add or Remove programs”.
From the “Add or Remove programs” dialog box, click the selection of “LineGene 9600” installation program and click “Remove” key.
Click “Remove” key and on the pop up “Add/Remove program” dialog box click “Yes”. The system will automatically uninstall the selected software.
When uninstalling program normally, log files of detection rusult are not deleted automatically. If customer confirms that they do not need these files, they may delete them automatically.
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LineGene9600 Start
Chapter 3 Start
Caution:
Before initial use of this instrument, correctly take out the fixing pin on its side and insert and tighten the lock into unlocker hole at its back; otherwise, the instrument would be unable to make fluorescence scanning (For removal of fixing pin, refer to chapter II, “3.1 Removal of fixing pin”).
1. Checks before start
After inserting the power plug and energizing this detection system, the followings shall be checked:
Check the voltage of power supply for consistency with system-required one.
Check the power wire plug for correct and reliable insertion to power socket.
Check the communication converter for correct insertion into the host and tightening and the lead plug for correct and reliable connection to the computer.
Check the update shift switch MODE of communication converter for being at normal state.
Check the environmental working environment and the storage condition for meeting requirements.
2. Start
In order to ensure effective communication connection between the instrument and the computer system, the system shall be started in the following sequences:
1 st step: Start computer display and host.
2 nd step: Turn on power switch on instrument’s rear bottom board.
3 rd step: Press Run Switch on front cover to set the system at running-allowable state.
4 th step: After entering operation system, start LineGene 9600 fluorescent quantitative detection system.
For starting software, click “LineGene96 20 0 /LineGene9640/LineGene9660/LineGene9680 ” from [Start]/ [program] menu or double click short-cut icon on desktop.
3. Starting software interface
Click one of the short-cut icons “LineGene 9620 / LineGene 9640 / LineGene 9660 / LineGene
9680" on desktop, the computer screen will show the following start image.
-10-
LineGene9600 Start
Fig. 3-1 Starting software interface
On the starting software interface, there are 4 buttons: Absolute Quantification, SNP, Relative
Quantification and Quit.
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LineGene9600 Start
Click “Absolute Quantification”, “SNP” and “Relative Quantification” button to make program enter corresponding detection program setup interface.
Click “Quit” button, close Starting software interface and exit program.
If you are still in one function module and you want to select another, it will not be available. For example, if you are running the absolute quantification, and you click "Relative Quantification", you will get a message as follows:
Fig. 3-2 Prompt dialog box
Note: Before doing experiment, please confirm first the computer hibernation state settings and hard drive state settings after opening the computer.
Setting method as follows: righ-click anywhere on your Desktop, select Properties, and click the Screen Saver, and then click the Power Button.
As below chart "Power Schemes", choose "Never" in the closed state of hard drive, and in hibernation, delete the hook before "Enable Hibernation" box.
Run the program and it will display the system window.
-12-
LineGene9600
The system window consists of the menu bar, the toolbar and the main form.
Start
-13-
LineGene9600 Absolute Quantification
Chapter 4 Absolute Quantification
1. Design Experiment
Start
This section describes how to design absolute quantitative experiment and covers new absolute quantitative experiment, inspection item setting, sample information setting, reaction plate setting and program setting.
Design experiment
Prepare for the reaction
Run the experiment
Experiment analysis
Experiment report
Data export
End
This chapter details the establishment, editing, saving, modification, running and analysis of absolute quantification detection program file of LineGene 9600 series
-14-
LineGene9600 Absolute Quantification fluorescent quantitative real-time PCR detection system
To build a new absolute quantification detection program file
Before newly building a detection program file, click “ ” to open program setup guide, which may give help in completing each setup before running.
The methods to build a new absolute quantification detection program file mainly include the followings:
After clicking “Absolute Quantification” button of “Starting software interface” (refer to Fig. 3-1), the system will automatically open a blank absolute quantification program setup interface (as shown in Fig. 4-1).
After clicking “ ” from “Setup Program” interface of detection file a blank “Setup Program” interface will occur.
Open “File (F) →new (N) Ctrl+N” to establishment a new blank “Setup Program” interface.
Open interface of blank absolute quantification detection file1.1 Create
New Absolute Quantitative Experiment
Click build New Experiment on Home interface and create absolute quantitative experiment window.
※
Absolute quantitative experiment can be also created by: a. Clicking FileNew on the menu bar b. Clicking New on the toolbar
-15-
LineGene9600 Absolute Quantification
1.2
Detector Setting
Click SetupDetector
Input experiment properties
Input the experiment name, user name and comment at the experiment properties column.
Detector Setting
Set up the Detector, Assay, Dye and Color.
※
If necessary, the user can also: a.
Add detector b.
Add assay c.
Delete detector d.
Delete assay e.
Add the detector in the Detector Library: click Add Detector Form Librarythe Detector
Library window will pop upselect the Detector on the window to be added
※
The user can also conduct Add, Modify and Delete operations in the item library.
-16-
LineGene9600 Absolute Quantification f.
Set up the detector, set up the assay, set up the dye name and set up the color
Set up reference dye
1.3 Sample Information Setting
Click SetupSample
Add sample information a.
Itemized addition: input ID in Sample ID press Enter add one piece of sample information b.
Batch addition: click Batch Addthe Batch Add window will pop up
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LineGene9600 Absolute Quantification
Delete sample information a.
Itemized deletion: select one sample click Delete delete the selected sample information b.
Delete all: click Clear Alldelete all sample information
Import/Export sample information a.
Click Import Sample Info the File Import window will pop up import sample information file in CSV format b.
Click Export Sample Infothe Save As window will pop upthe sample information will be exported in CSV file format
Set up sample information
1.4
Reaction Plate Setting
Click SetupPlate
Set up the inspection items of reaction plate a. Select reaction plate hole site: click Reaction Plate Hole Site
Apart from reaction plate hole site, the user can also right click the reaction plate hole site to Copy, Paste and Add New Detector. Adding new detector will open the Edit Detector
Library window.
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LineGene9600 Absolute Quantification b. Select Assay item and modify the property, concentration and concentration unit.
Property Name Concentration
Concentration unit
Unknown
Standard
Negative
Positive
NO
YES
NO
NO
Copies/ml
IU/ml
Fg/ml
Pg/ml c. Select a sample and the list displayed will change d. Zoom In, Zoom Out and reset the reaction plate.
e. Sample Auto Arrange f. Check Well Table
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LineGene9600 Absolute Quantification
1.5 Program Setting
Click SetupProgram
Run Programs Setup a. Create new stage: the user can create new Hold Stage, Cycling Stage or Melting Stage
※
The user can also click Add Stage directly and the default will be creating new Cycling
Stage.
b. Create new step: the user can create new step Before or After the currently selected step
※
The user can also click Add Step and the default will be adding new Step at the end of currently selected stage or after currently selected step.
c. Delete: the user can delete currently selected step or stage d. Display form: click Display With Table new window will pop upthe details of current experiment will be displayed in table e. Set up the experimental data of the hold stage, cycling stage and melting stage melting section f. Set up hotlid temperature and liquid quantity
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LineGene9600 Absolute Quantification
2. Prepare for Reaction
Start
The user should make full preparations prior to the experiment:
• Ensure appropriate materials are used.
• Ensure the arrangement of PCR reaction plate is consistent with the setting layout of reaction plate in Section 1.4.
Design experiment
Prepare for the reaction
Run the experiment
Experiment analysis
Experiment report
Data export
End
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LineGene9600 Absolute Quantification
To open and modify absolute quantification detection program files
To open an absolute quantification program file
The methods to open parameter setup file of absolute quantification real-time PCR detection program include the followings:
After entering “Setup Program” interface, click “ ” or “File(F)→Open (O) Ctrl+O”, pop up a dialog box, find out directory of the target file, select target file, then click “Open (O)” to open a setup parameter file of an edited system default directory.
Click “File (F) →Import Setup Parameters (I)” to open saving setup parameters in the system default directory (impossible to import running result).
Click “System Parameters (P) → Load Customized Parameters (P)” to open the saved user customized parameter file of file format *.ucp.
The methods to open the detection result file of absolute quantification PCR detection program include the followings:
Through “File (F) → Import(M)” open the absolute quantification detection result file of non-system default directory (detection program file format *.fqd,*.fqj).
Through “File (F) →Import SNP File (N)” open SNP detection result file of non-system default directory (detection program file format *.fqs,*.fqd).
Through “File (F) →Import Relative File (R)” open the relative quantification detection result file of non-system default directory (detection program file format *.fqx).
The opened interface simultaneously displays the test name and modification time, which will provide information of helpful note for following data analysis or opening program file.
To open a program file interface
Caution
:
Under opening mode, the user can input set parameters of detection program but cannot input detection result.
The file requiring analysis of the detection result may be opened only through
“Import”, “Import SNP File” and “Import Relative File”.
To modify an absolute quantification program file
Opening a not-running detection program file may modify each setup parameter of program file.
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LineGene9600 Absolute Quantification
Through opening a runned detection program file, the parameters of “Setup program” interface can’t be modified.
For saving the modified file, refer to “3. Saving of absolute quantification detection program file”.
Storage of absolute quantification detection program file
After program running the setup parameters and detection result of the absolute quantification detection program file will be automatically saved in system default directory and the detection file also may be as per actual requirements subjected to manual customized saving.
Click “ ” or open “File (F) →Save (S)” to save the file in default directory.
Click “File (F)→Save As (A)” to pop up file save as dialog box, under the system default directory select saving sub-directory and after determining the saving name click “Save”.
Click “File (F)” and press “Ctrl+S” to make rapid saving operation.
Click “File (F) →Export (X)” to save the detection file in non-system default directory with file format *.fqj.
Click “Saved User Customized Parameters (P)” of “System Parameters (P)” pull-down menu and save the setup parameters as user customized parameter file with file format *.ucp.
When saving detection program file, the default saving folder is user name and the default saving file is the test name. In case of no user name or test name, the default saving folder is default and the default saving file hh(hour)-mm(minute)-ss(second)-mm(month)-dd (day)-yyyy(year).
name is: fqd
Program file saved-as interface
To set up detection program
Compositions of detection program
The absolute quantification real-time PCR detection program file consists of parameter-setup, running and detection result of the detection program. The entire detection system consists of Setup Programs, Sample Data, Run program, Quantitative Analysis, Melting Curve
Analysis and Data Inquiry.
Setup Programs: This part mainly sets up the detection program, program section and its
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LineGene9600 Absolute Quantification parameters.
Sample Data: It sets up the sample information of detection program.
Run program: During program running process temporarily modify the setting up of the program, and real-time observe the change of each detection parameter of the detection program.
Quantitative Analysis: It makes quantitative analysis of detection result and saves and outputs the quantitative analysis result.
Melting Curve Analysis: It makes melting curve analysis of detection result and saves and outputs the melting curve analysis result.
Data Inquiry: Find out the sample information and its detection result of the corresponding sample as per the input inquiry condition.
Absolute quantification detection program setup interface
Procedures of setting-up detection program
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LineGene9600 Absolute Quantification
To input user name
The user can input the user’s name in the setup program interface directly.
Since at utmost 20 characters are shown in the user name column, for the input name of more than 20 characters merely the first 20 ones are displayed.
The user name does not input to the user name column and thus the edited program file is saved in the default folder.
When saving program file the user name will be automatically established as the default folder name.
To input test name
The user can input test name in the setup programs interface directly.
Since at utmost 20 characters are shown in the test name column, for the input name of more than 20 characters merely the first 20 ones are displayed.
Setting-up of hot-lid
Click selection box at left of hot-lid to select opening hot-lid or not.
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LineGene9600 Absolute Quantification
After selection, directly input the setting up temperature of hot-lid.
The temperature set for the hot-lid shall be at least 10°C higher than the maximum temperature during sample running to avoid reagent evaporation.
The setting up temperature range for the hot-lid is 30°C~110°C.
Setting-up of liquid quant
The user can input liquid volume in the setup programs interface directly.
The setting up range of liquid volume is 5μl~100μl and it may be setup as per actual conditions to optimize the system’s detection control.
Setting-up of analysis mode
The analysis mode can be determined as per test detection purpose or contents: “Cycle/Temp.” or “Melting”. From the setting up program interface click “Cycle/Temp.” or “Melting” button to select corresponding analysis mode and setup the analysis mode.
Analysis mode setup
Setting-up of cycles/constant temperature analysis mode
After clicking “Cycle/Temp.” button from the setting up program interface, the program segment and program section setup column will automatically pop up the added “Cycle/Temp. 1” program and program section.
Each program segment may be edited through adding, deleting or moving in the program segment column.
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LineGene9600 Absolute Quantification
In the program section, it can be edited, such as quantity, target temperature, time, rate, 2nd temp., step size, step delay, grad temp., sample mode and cycles.
a) Adding /removing /moving main program segment
Adding: After clicking “Cycle/Temp.” button, the system will automatically in the right program editing column add program 1 “Cycle/Temp. 1”. Re-clicking “Cycle/Temp.” will add program
2, 3…shown in left of Fig. 4-6). For remark or explanation to program, click remark column of the said program to directly input the information of 20 characters.
Removing: Select the to-delete program item, click the right key, and select “Remove” to delete the unnecessary program segment.
Moving: Select the program item that need to be moved, click the right key, and select “Moveup” or “Movedown” to adjust the position of program segment.
Setup interface of cycles/constant temperature main program segment
b) Adding /deleting program section
Adding: After clicking “Add” button, a newly added program section will occur following the last program section.
Deleting: Click any position of the line of the program section to select the said program section and then click “Delete”.
Inserting: After selecting a program section with mouse and clicking “Insert”, a newly inserted program section will occur over the said program section.
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LineGene9600 Absolute Quantification
Setup interface of cycles/constant temperature program section
Note:
The deleting /adding of program section may be used only with “Cycle/Temp.” analysis mode.
The program section is inserted only before the current program section and in case of mistake in operation it may be re-inserted after deleting.
Each program segment may setup at utmost 20 temperature control program sections.
Caution:
The parameters of newly-added program section are system’s default parameters and they may be modified as per requirements.
c) Setting up cycles of detection program
The setting up of cycles is merely applicable to the program segment of analysis mode
“Cycle/Temp.” and in case of analysis mode “Melting” the program mode display interface will have no setup column of cycles.
The set value of cycles may be directly input.
The input value of cycles is integral from 1 to 99.
Setup of cycles
d) Setup of parameters of program section
Target temperature: It may be directly input as per actual condition with setting up range of
4~105°C.
Time: It may be directly input.
After clicking and selecting time column, pull-up and pull-down key will appear at its right side
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LineGene9600 Absolute Quantification and time may also be input through clicking pull-up and pull-down key.
The time display format of time column is hour: minute: second.
Rate: It may be directly input and its default value is 4.0°C/s and setting up range is 0.1°C/s~
4.0°C/s.
2nd Temp.: It may be directly input and its default value is “-” and setting up range is 4°C~
105°C.
Step Size: It may be directly input and its default value is “-” and setting up range is 0.1°C~
9.9°C.
Step Delay: It may be directly input and its default value is “ - ”, the minimum is 1 and the maximum can’t be higher than the set cycles.
Grad Temp.: It may be directly input and its default value is “-” and setting up range is 1°C~
36°C.
Setup of parameters of program section
The time and target temperature of each program also may be modified in analog display area.
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LineGene9600 Absolute Quantification
As shown in the below diagram, the top of curve is the target temperature and the active figures are clicked to modify the start temperature and the temperature value may be directly input. The bottom of the curve is time and the active figures are clicked to pop up up/down key and to modify time.
Modification of each program time and temperature of analog display area
e) Setup of sample mode
Merely a temperature control program section is selected for sampling through the side pull-down box. If the temperature control program section is selected as “Single” sampling, after completion of running of this section, the system will scan and collect fluorescence data of the reagent sample.
Setup of melting curve analysis mode
Setting-up of melting curve analysis mode and “Cycle/Temp.” are basically same merely with different parameter contents of program section. In the program section, the user can edit the target temperature, start temperature, time, step temperature and sample mode.
a) Setup of parameters of program section
Target: It may be directly input as per actual condition with setting up range 4~105°C.
Start temperature: It may be directly input as per actual condition with setting up range 4 ~
105°C.
Time: It may be directly input.
After clicking and selecting time column, pull-up and pull-down key will appear at its right side and time may also be input through clicking pull-up and pull-down key.
The time display format of time column is hour: minute: second.
Step: It may be directly input and its default value is “-” and setting up range is 0.1°C~9.9°C.
Caution: The
step temperature denotes the each ascending or descending temperature value.
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LineGene9600 Absolute Quantification
Setup interface of melting curve program section
b) Setup of sample mode
“None” or “Step” is selected through the pull-down box of sample mode column. If “None” is selected, during the detection program running process no sampling shall be made. If
“Step” is selected, during the detection program running process for each increase of a step temperature one sampling shall be made.
Target temperature: 95°C
Time: 20s
Step temperature: 0.5°C
Start temperature: 60°C
Sampling point:
Illustration of setup of melting curve
Setting-up of end-running mode
After clicking “End” button from the setting up program interface, the program segment and program section setup column will automatically pop up the added “ End” program and program section.
In the running program segment setup column, “ End” program is merely located at the last of each program segment and it may be deleted.
In the program section, its target temperature and rate may be edited.
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LineGene9600 Absolute Quantification
Setup interface of end-running mode
a) Adding /removing /moving main program segment
Adding: After clicking “ End” button, the system will automatically in the right program editing column add termination routine “ End” and the termination routine is merely allowed to add once. For remark or explanation to program, click remark column of the said program to directly input the information of 20 characters.
Removing: Select the “Remove” program item, and click the right key.
Moving: “ End” is unable to execute the moving command and the said program segment is merely located at the last of each program segment.
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LineGene9600 Absolute Quantification
Setup interface of main program segment
b) Setup of parameters of program section
Target: For “SOAK” mode, the system default target temperature is 8°C.
For non “SOAK” mode, the target temperature may be directly input with setting up range 8~
105°C.
Rate: It may be directly input as per actual condition with setting up range 0.1~4°C/s.
Setup interface of program section
Caution:
If “SOAK” mode is selected in termination routine segment, at end of program running it will execute the below 10°C low-temperature storage command.
Input of sample information
Click “Sample Data” to enter sample information setup interface and set up the input, saving and modification of sample information.
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LineGene9600 Absolute Quantification
Setup interface of sample information
Setting-up of detection test tube display area
In the detection test tube display area, the detection test tube may be selected and the sample information of the selected test tube may be edited through the following modes:
Clicking “ ” of test tube display area to select single test tube.
Through pressing Ctrl key several test tubes may be irregularly selected.
Through pressing Shift key several test tubes may be regularly selected, i.e. the selected test tubes are arranged in a rectangular. If pressing-on Shift key and selecting “A1” and then
“F9”, all test tubes in the rectangular area with “A1” and “F9” as diagonal will be selected
Through pressing-on and moving mouse left key, several test tubes may be regularly selected.
Clicking line number “A, B, C…” of the table, the entire line of test tubes may be selected.
Clicking row number “1, 2, 3…”of the table, the entire row of test tubes may be selected.
After selection of detection test tube, the sample information of the selected detection test tube
(except for sample color) may be integrally input and modified. For example, if “A1” and
“B2” are selected and “A1” sample name is changed as “Sample 1” and “B2” sample name will be correspondingly changed as “Sample 1”.
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LineGene9600 Absolute Quantification
Irregular selection of test tube Regular selection of test tube
Selection of entire-row test tubes Selection of entire-line test tubes
Selection of test tube display
Group setup
In the group setup area, click the pull-down arrow at right of group number and from the pull-down box select the group numbers of sample: 1,2,3,4 and click pull-down arrow at right of the groups to setup the group of the selected sample.
Group and concentration setup interface
Input of concentration
In the concentration setup area, click pull-down key at concentration unit right and from the pull-down box select dye concentration unit: copies/ml, IU/ml, fg/ml and pg/ml. In the concentration setup area current dye concentration of the selected sample may be directly input.
Dye addition
The dye may be added as follows:
In dye addition selection area, after clicking dye and then “Add (A)” button, the dye will be automatically identified to the detection test tube and the right side dye attribute column and the detection test tube display area will also display corresponding color.
Click “Delete (D)” button to cancel corresponding dye setup.
Click “Undo (U)” button to cancel the latest operation.
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LineGene9600 Absolute Quantification
Click “Redo (R)” button to cancel the undo operation.
In the dye attribute setup area; click the corresponding position of table to select the sample attribute.
At selection of dye, when a dye is selected, partial dyes display grey, which denotes that these dyes are in a same channel and they are unnecessary to re-select.
The sample will have the following attributes: standard, unknown, negative, positive and blank.
Setup of dye addition and attribute
Editing of sample information
In the sample information area, the sample materials information has totally 22 default items, of which the group, concentration, dye and attribute shall be setup in corresponding setup column and the others may be selected and input as per actual requirements.
The position of sample column in the list may be modified through mouse drag.
Sample information input interface
The sample materials are input as follows:
Well: Click “+” to develop the tabulation and list contents of 8 lines “F1~F8” and show the dye information of 8 detection channels corresponding to the said well sample.
Sample: After clicking a column, the contents may be directly input. The said column can’t be complete blank; otherwise the program will be impossible to automatically run. The default sample name is well number, e.g. “H12”.
Color: Double clicking a column will pop up a color dialog box. After selection of a new sample color in the color dialog box, clicking “Enter” will replace the sample default color.
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LineGene9600 Absolute Quantification
Color dialog box
Type: The contents may be directly input.
Sample No.: The contents may be directly input.
Number: The contents may be directly input.
Name: The contents may be directly input.
Sex: Click and select a column, double click it to pop up the pull-down key at its right, click the pull-down key and from the pull-down box select male, female and unknown.
Age: The contents may be directly input.
Outpatient No.: The contents may be directly input.
Inpatient No.: The contents may be directly input.
Bed No.: The contents may be directly input.
Case No.: The contents may be directly input.
Doctor: The contents may be directly input.
Dept.: The contents may be directly input.
Submitting date: Click and select a column, double click it to pop up the pull-down key at its right, click pull-down key, pop up calendar, move the mouse to the selected date on the calendar and click the said date to input detection date.
Diagnosis: The contents may be directly input.
Sampling time: Click and select a column, double click it to pop up the pull-down key at its right, click pull-down key, pop up calendar, move the mouse to the selected date on the calendar and click the said date to input sampling time.
Status: The contents may be directly input.
Nationality: The contents may be directly input.
Notes: The contents may be directly input.
There are 3 shortcut keys below the sample information list: Clear (E), default (D) and restore
(R).
Clear (E): Clicking the said key will eliminate all sample materials.
Default (D): Clicking the said key will automatically show the setup of system default sample materials.
Restore (R): Clicking the said key will restore the sample materials to its original state.
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LineGene9600 Absolute Quantification
Selection of sample data column
If the setup of system sample data column is deemed unsuitable, the sample data column may be selected. Click “Sample Data (S)” in menu column, from the pull-down menu select
“Select Sample Data Column (E)”, pop up sample data columns and select dialog box.
Dialog box of sample data columns selection
Click “Add (A)”or “Remove (D)” to select desired column, which will be displayed in the right box.
Select a sample column in the right box and click “Move up (U)”or “Move down (D)” to change the position of the said column in sample information list.
Click “Add All (W)” or “Remove All (M)” to make integral adding or deleting operation.
Customized sample data columns
Customized sample data columns may be added if necessary. From the pull-down menu of
“Sample Data(S)” select “Customized Sample Data Column … (C)” and pop up dialog box of customized sample data columns.
Dialog box of customized sample data column
Click “Add (A)” button to automatically add one line. Click column 1 to make edition and input the name of customized sample data column.
After clicking “OK (O)” button, the customized sample data column will appear in the column at left of dialog box of the sample information selection column.
Clicking “Remove(R)” button will delete the customized information column.
Clicking “Cancel (C)” button will close the dialog box.
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LineGene9600 Absolute Quantification
Calculation of grad temp.
At setup of gradient parameter, the temperature of each sample well at each set gradient state may be referred to grad temp. calculation diagram. The sample is divided into 12 rows and each row has the same grad temp.. Clicking the “Grad Temp. Calculation (C)…” of the
“System Parameters (P)” pull-down menu will pop up a calculation grad temp. dialog box.
Grad temp. calculation diagram
Gain setup
The LineGene 9600 series Fluorescent Quantitative PCR Detection System has totally 8 detection channels F1, F2, F3, F4, F5, F6, F7 and F8, each channel’s gain can be adjusted independently.
Gain is the amplification factor of fluorescence intensity signal and adjustment of gain value will change the magnitude of fluorescence intensity output signal, scope of adjustment is from
0 to 20. The system provides default gain value to greatly shorten the time to determine the gain value through test and thus greatly enhance instrument suitability. Man-made setup may be made as follows:
Clicking “System Parameters (P) →Gain Setup (G)…” menu enter “Gain Setup”.
Directly input gain value and click “OK”.
Setup gain
Function of best gain
Because gain set by user is improper, scanned fluorescence value may probably too large or too small. If fluorescence value is too high, electrical signal will saturate, which causes display of interface corresponds with actural amplification. If fluorescence value is too small,
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LineGene9600 Absolute Quantification it is not easy to observe. The software takes amplified reagent as sample. Instrument adjust again value automatically during scanning, so that object value fluorescence gets best effect. (Fluorescence value of aplification apporaches 9000)After measurement, the gain value gotten from scan is the best gain value. User may invoke best gain at any time.
Best gain of every instrument has been measured by using usual reagent at ex-factory. When user’s reagent is different from dye name of channel list or the reagent is especial, user should measure best gain.
The following is calibration procedure:
User may select reagent,which is single character and pure dye, of one or more channel to measure at the same time, and put reagent in appointed wells.
Channel
F1
F6
F7
F8
F2
F3
F4
F5
Reference well of best gain reagent
A1
A1
A2
A3
A4
A5
A6
A7
Caution: Because F1 and F2 are in the same well, dyes of F1,F2 must measured respectively.
b)In the interface of sample data, to select dye, then select the “Best Gain Measurement” of the
“System Parameters” pull-down menu.
c)To input password“bioerpcr”in the following window.
d)To click “Yes” in the following window, system measures best gain.
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LineGene9600 Absolute Quantification e) After running, file “Gain******.coe” is saved in installation folder, thereinto instrument number is correspondence with *. User can see dye in “Best Gain Dyes Measured(M)…” of
“System Parameters (P)” pull-down menu.
f) Clicking “System Parameters (P) →Gain Setup (G)…” menu enter “Gain Setup”.
g) To select “Best Gain” from “Gain Setup” interface, then click button “run” after setting parameters. During detection program running, best gain value of dye of each channel will be imported. If best gain value of some dye is not acquired, pop the prompt dialog box as follows:
Caution: When amplification, best again or less than best gain is adopted.
Addition of customized dye
If the detection-required fluorescence dye is not in the system default fluorescence dye, corresponding fluorescence dye may be added in the system through “Add Customized
Dyes”.
Click the “Add Customized Dyes (Y)…” of “System Parameters (P)” pull-down menu to pop up
“Add Customized Dyes” dialog box.
Directly input the new-added dye name to the “Dye Name” column
Directly input the new-added dye detection channel to “Dye Channel”.
Click “OK” to save the new-added dye to the pull-down menu of “Analysis Options”.
Select “Clear Customized Dyes” and click “OK” to delete all customized dyes from pull-down menu of “Analysis Options”.
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LineGene9600 Absolute Quantification
Module scanning mode
Addition of customized dyes (e.g. F9)
In LineGene 9600 detection process, there are 2 fluorescence scanning modes (entire scanning and in-line scanning), which may be setup through “Block Scan Method”
Click “Select Block Scan Method (M)…” of “System Parameters (P)” pull-down menu to pop up
“Block Scan Method” dialog box.
After clicking and selecting “Whole Block Scan” and clicking “OK”, the instrument will conduct entire scanning during running.
Re-clicking “Whole Block Scan” will cancel the selected “Whole Block Scan” and activate “Line
Scan Setup”. “Line Scan Setup” box has 8 options (A, B, C, D, H) and through clicking 8 options, the in-line scanning may be selected and setup. After clicking “OK”, during running the instrument will make in-line scanning as per the setup.
Setup of select block scan method
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LineGene9600 Absolute Quantification
To copy baseline parameters to install directory
Click “Copy Baseline Parameters to Install Directory (L)…” of “System Parameters (P)” pull-down menu to pop up “Open” dialog box. You may open baseline parameters file, format of file: Baseline******.b96, thereinto instrument number is correspondence with *.
Copy Baseline Parameters to Install Directory
Baseline parameters file “Baseline******.b96” has saved in disc at ex-factory.
If instrument serial number does not match baseline parameters under instrument install directory, it will pop up dialog box, click “Yes”, copy parameters according to prompt, run program after quitting. If instrument serial number matches baseline parameters, system does not prompt when running.
Dialog Box of Prompt
To copy crosstalk parameters to install directory
Click “Copy crosstalk Parameters to Install Directory (S)…” of “System Parameters (P)” pull-down menu to pop up “Open” dialog box. You may open crosstalk parameters file, format of file: Scan******.coe, thereinto instrument number is correspondence with *.
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LineGene9600 Absolute Quantification
Copy Crosstalk Parameters to Install Directory
Crosstalk parameters file has saved in disc at ex-factory, File that is made by user’s calibration may be selected under install directory or directory that user appoints.
After success of copy, you must quit program, then open it.
To copy best gain parameters to install directory
Click “Copy Best Gain Parameters to Install Directory (B)…” of “System Parameters (P)” pull-down menu to pop up “Open” dialog box. You may open best gain parameters file, format of file:
Gain******.bgn, thereinto instrument number is correspondence with *.
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LineGene9600 Absolute Quantification
Best gain parameters file has saved in disc at ex-factory, File that is made by user’s calibration may be selected under install directory or directory that user appoints.
After success of copy, you must quit program, then open it.
To copy fluorescence calibration parameters to install directory
Click “Copy Fluorescence Calibration Parameters to Install Directory (B)…” of “System Parameters
(P)” pull-down menu to pop up “Open” dialog box. You may open fluorescence calibration parameters file, format of file: flr******.prp, thereinto instrument number is correspondence with
*.
Fluorescence calibration parameters file has saved in disc at ex-factory, File that is made by user’s calibration may be selected under install directory or directory that user appoints.
After success of copy, you must quit program, then open it.
Caution: User can measure crosstalk dyes after fluorescence calibration.
System prompt and alarm
The system prompt includes curve prompt function and buzzer alarm function. From the
“Options for System Alarm and Tips” dialog box, the system prompt and alarm may be setup.
After starting “Tips for Curves“, when the mouse point moves to curve, which is overstriking ,information prompt box of the curve will appear and the said information box will display related information of the mouse-located curve (detection sample well, X coordinate, Y coordinate, selected dye, and groups of detection sample).
After starting “alarm”, in case of instrument troubles or end of run, the instrument will send buzzing sound.
Click “Options for System Alarm and Tips (O)…” of “System Parameters (P)” pull-down menu to pop up “Options” dialog box.
After clicking “Tips for Curves”, when “” of the current page changes to “” it will denote that the said function is selected.
After clicking “alarm”, when “” of the current page changes to “” it will denote that the said function is selected.
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LineGene9600
Directly input buzzer alarm duration into “Alarm Duration (s)” column.
Click “OK” to save the system prompt setup.
Absolute Quantification
Setup and display of curve prompt function
Load customized parameters
At detection of repeated items, in order to decrease repeated input the saved customized parameters may be called through “Load Customized Parameters”.
Click “Load Customized Parameters (P)…” of “System Parameters (P)” pull-down menu to pop up “Password for System Parameter Setup” dialog box.
Input system password and click “OK” to pop up “Open“ File dialog box.
Select the desired file and click “Open” to open the saved parameters.
User customized parameter file format: *.ucp
To open user customized parameter file
Saved as user customized parameters
Through the “Save Customized Parameters” the seted detection program parameters may be saved for call in next detection.
Click the “Save Customized Parameters (P)…” of “System Parameters (P)” pull-down menu to pop up “Password Setup” dialog box.
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LineGene9600 Absolute Quantification
Input and confirm password (Caution: password over 3-digit) and click “OK” to pop up “Save As” dialog box.
From “File Name” column directly input file name and click “Save” to save the customized parameter file.
File saving type: customized parameter files (*.ucp).
To save user customized parameter file
Crosstalk measurement
Any dye has a wide light spectrum. Filter of adjacent wavelength of F1-F8 is effective, and it is normal physics for them to overlap.
Usually, this software can correct the fluorescence among channels. It records every channel to deduct this impact later, minimize the “interference” among channels and avoid wrong judgment in same-tube multi-color synchronous detection.
In order to eliminate effect which occurs between one channel dye and the others,so that user can get multicolor detection results, the software adds function of crosstalk measurement
F1’s impact on F2:
Crosstalk effect
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LineGene9600 Absolute Quantification
After calibration, the impact disappears:
Start
Design experiment
Prepare for the reaction
Run the experiment
Experiment analysis
Experiment report
Data export
Crosstalk of every instrument has been measured by using usual reagent at ex-factory. User may measure gain reagent. If there is no reagent in list of dye, user should name dye at suitable channel, then measure crosstalk.
The following is calibration procedure: a) User may select reagent,which is single character and pure dye, of one or more channel to measure at the same time, and put them in appointed wells.
Channel
F1
F2
F3
F4
F5
F6
F7
F8
Pure water
3. Run the Experiment
Reference well of crosstalk reagent
A2
B2
C2
D2
E2
F2
G2
H2
A3
This section describes how to run operate the experiment after loading the reaction plate and covers the operating of florescence curve, the operating of temperature curve and prog
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End
LineGene9600 Absolute Quantification
Caution:
Before start, make checks and correctly start the system.
The green lamp of run switch shall light on and the system pushed to send locking sound and the alarm lamp is not on.
If the module is pulled out or is improperly pushed, the b)To run a program whose sample mode is none, and the reagent volume software will show prompt and the alarm lamp will light on.
At this case, temperature program can run but the fluorescence scanning data would become invalid.
is 40μl.
During running, to select “Crosstalk Measurement ” of “System Parameters” pull-down menu.
c) To input dye and suitable gain for channel in the following dialog box, and select “Measure
Crosstalk Coefficient”, then click “ok”, crosstalk coefficient is measured.
After running, file “scan******.coe” is saved in installation folder, thereinto instrument number is correspondence with *. In interface of run, pop up the upper window after run, user may select
“Cancel” to quit measure, and may resume the upper produce to measure. Again(In order to assure correct data, crosstalk is required to measure twice) User can see dye in “Crosstalk
Dyes Measured (T)…” of “System Parameters (P)” pull-down menu.,during checking, please confirm that Default.ser is in installation directory.
Caution: To selected dye name does not correspond with actual dye, which causes wrong data.
Because spectrum overlap of dye is large between channel F1 and channel F2(such as hex, tamra), crosstalk does not measure. Generally, user use one from F2,F3.
User may select reagent,which is single character and pure dye, of one or more channel to measure at the same time, and may add and cover, the saved file name: gain******.coe and scan******.coe, thereinto instrument number is correspondence with *. When measuring, operate software according to prompt, user may select dye name or define new dye name, set
“gain” to fit value of channel when running.
The reagent volume is 40μl, and it takes about 45min 85min during measure.
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LineGene9600 Absolute Quantification
Flourescence Calibration
To open amplification result of FAM which includes the same concentration. Because of heterogenicity of fluorescence detection among wells, discreteness of CT value is obvious.
There are discrepancy in baseline of different wells and in actual fluorecent gain. “Fluorescence
Calibration” is to remedy heterogenicity of fluorescence increment.
After uniformity calibration of fluorescence, discreteness of CT value is improved obviously.
The following is calibration procedure:
In interface of quantitative analysis, to import analysis result, and to select the “Fluorescence
Calibration” of the “System Parameters” pull-down menu.
Caution: To delete file “*.prp” from installation directory before calibration.
To input password“bioerpcr”in the following window.
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LineGene9600 Absolute Quantification
To input the cycle start in the following window (To select train cycle of fluorescence rise point, generally,select cycle 1-3 after rise point), and select “Need Fluorescence Calibration”, then click “OK”. System bases on fluorescence data of three sampling pionts of every well which is from start cycle, and caculate calibration coefficient automatically, then calibrate fluorescence uniformity of the result. Calibration coefficient of every well is saved as default file. When openning new file of result(The result must correspond with serial number of instrument), system can calibrate automatically according to saved calibration parameter, then discreteness of CT is improved substantially.
Causion: In order to get better result, user may calibrate again under the said situation.
Do not continuously calibrate different cycle points when file “*.prp” has made.
File information
After opening a result, click “File info. (N)…” of “System Parameters (P)” pull-down menu to pop up dialog box, in which display the file name from the data file, and the file name is with instrument number and all modification parameters.
If there is no opening file, “File info.” is under unusable state.
Less common items of system parameters
In “System Parameters (P)” pull-down menu, there are items that are used at ex-factory, as follows: select system parameters, Y axis calibration, baseline measurement, crosstalk measurement, crosstalk dyes measured.
In “Select System Parameters”, there are X origin calibration, measure gain ratio and crosstalk coefficient, clear run time, write machine serial NO. , read machine serial NO., write PMT parameters, read PMT parameters. These items can be used if instrument is running. In
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LineGene9600 Absolute Quantification addition, if you want to use these functions, you must enter password, Password has been set at ex-factory, you may ask manufacturer for password directly.
“Y Axis Calibration” means to calibrate Y axis automatically, This item can be used if instrument is running. In addition, if you want to use this function, you must enter password, Password has been set at ex-factory, you may ask manufacturer for password directly.
“Baseline measurement” includes two items: baseline measurement, substract baseline. If selecting baseline measurement, you may measure baseline of instrument automatically, and calibrate baseline automatically, results will be saved automatically in file “baseline******.b96” under install directory. If selecting substract baseline, system will carry out order of cutting baseline value in baseline parameter file “baseline******.b96” when fluorescence curve appers.
Otherwise, system will not operate cancelling background. This item can be used if instrument is running. In addition, if you want to use this function, you must enter password, Password has been set at ex-factory, you may ask manufacturer for password directly.
“Insert Cal. Para”, “Export Raw Data” and “Export Cal. Para” are used to deal with data by company, and user need not use these function.
System Parameters pull-down menu
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LineGene9600 Absolute Quantification
Running of detection program file
Caution:
Before start, make checks and correctly start the system.
The green lamp of run switch shall light on and the system shall be at running state.
Caution:
Before running of program, the module shall be smoothly pushed to send locking sound and the alarm lamp is not on.
If the module is pulled out or is improperly pushed, the software will show prompt and the alarm lamp will light on.
At this case, temperature program can run but the fluorescence scanning data would become invalid.
3.1
Preparation for reagent sample
Preparation for reagent: LineGene 9600 series fluorescent quantitative PCR detection system adopts 0.2ml centrifugal test tube to place reagent sample and 10μl ~ 50μl is suggested for sample optimal reaction system.
Optic lucid tube on bottom such as normative single valve, calandria, full apron plate, and so on, are allowed in strument.
Centrifugal operation: Before placing into the instrument, the test tube with reagent sample shall be subjected to centrifugal operation to ensure that the reagent is at test tube bottom and the inside is free of bubble.
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LineGene9600 Absolute Quantification
To insert test tube: If the sample quantity is less than well quantity of module , the sample test tubes shall be evenly distributed in module well as far as possible, so as to make the hot-lid steadily compress on the top of test tube during running. The load of module shall be even so as to ensure even and consistent temperature change of each test tube.
Correct
The sample is at reaction test tube bottom
Incorrect
The sample test tube requires sufficient speed
At centrifugal operation it requires sufficient centrifugal time
Correct position of sample in test tube
3.2 Start Run
Click Start Run
※ The default running interface is florescence curve
Operating confirmation a. Modify hotlid temperature and liquid quantity b. Gain parameter setting c. Target florescence value setting
※
If necessary, the user can modify above parameters directly in this window.
After it starts running, the user can: a. Skip the current stage b. Plus cycle
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LineGene9600 c. Minus cycle d. Stop run
Plot display setting a. Assay item b. Plot color
3.3 Monitor Operation
Monitor florescence curve: click Florescence Curve
Absolute Quantification
Monitor temperature curve: click Temperature Curve
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LineGene9600 Absolute Quantification
3.4 Program Setting
The user can only check the program setting but cannot make modification.
Time-temperature display during running 3.5
Working state indication lamps on instrument
The panel at upper left of instrument is fixed with 3 state indication lamps and they are related with system state during running of program:
Running: The indication lamp lights green, which denotes that the entire machine is under program running state.
Alarm: The indication lamp lights yellow, which denotes that the module is not fully pushed in and the hook is not fastened.
Error: The indication lamp lights red, which denotes that the instrument is in troubles.
Caution:
For prolonged shutdown after end of program running, set the power switch at instrument’s rear bottom board at “OFF” and cut off power supply. After turning on power, the hot-lid and module will run as per default state.
The front cover of instrument is fixed with a self-locking key to control energizing of its inside control system:
Run Switch: running/standby switch.
After pressing this key, the green loop lamp lights on instrument inside system is energized and the user is allowed to run the program.
After pressing this key again, the key will spring out, the green loop lamp lights off, instrument inside system is de-energized and the system is under standby.
Caution:
The run switch is for user’s easy operation and is merely used for temporary or
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LineGene9600 Absolute Quantification short time closing control system. When the system is under standby state, the instrument inside AC circuit is live.
3.4.1
Prompts likely occurring during running
Hot-lid temperature sensor alarm prompt
Sink temperature sensor alarm prompt
Environmental temperature sensor alarm prompt
Module temperature sensor alarm prompt
Module sensor short-circuit or short-circuit alarm prompt
Caution:
In case of temperature alarm display during running of program, the PCR detection system running program shall be terminated, the instrument shall be switched off and then re-started.
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LineGene9600 Absolute Quantification
4. Experiment Analysis
Start
This section describes how to view the experiment analysis result after the operating experiment and adjust parameter reanalysis. This section covers the analysis of amplification curve and standard curve, adjusting parameter reanalysis and parameter importing.
Designed experiment
Prepare for the reaction
Run the experiment
Experiment analysis
Experiment report
Data export
End
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LineGene9600
4.1 Check Results
4.1.1 Check the Amplification Plot
Click Analysis Amplification Plot
Absolute Quantification
Check the amplification curve a. Set up color b. Set up plottype c. Set up show dye
※
When the background color of a dye name is blue, it means to be displayed; while white indicates not to be displayed.
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LineGene9600 Absolute Quantification
Check the reaction plate a. Select relation plate hole site and check corresponding hole site curve
※
The default is all hole sites are selected b.Zoom In, Zoom Out and Reset the reaction plate c. Check well table information d. Check results summary
Set up assay a. Set up Assay b. Set up Threshold c. Set up automatic baseline
※ When the threshold value is not automatic, the user cannot set up the automatic baseline
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LineGene9600
4.1.2 Check Standard Curve
Click Analysis Standard Curve
Check standard curve a. Set up assay
Absolute Quantification
Check the reaction plate a. Select relation plate hole site and check corresponding hole site curve
※
The default is all hole sites are selected b.Zoom In, Zoom Out and Reset the reaction plate c. Check well table information d. Check results summary
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LineGene9600 Absolute Quantification
4.1.3 Check Melting Curve
Click Analysis Melting Curve
Check the melting curve a. Check the fluorescence curve b. Check the derivative curve c. Set up color
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LineGene9600 Absolute Quantification
Check the reaction plate a. Select relation plate hole site and check corresponding hole site curve
※ The default is all hole sites are selected b. Zoom in, zoom out and reset the reaction plate c. Check well table information
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LineGene9600 Absolute Quantification
Set up assay a. Set up assay b. Set up color
4.2 Adjust Parameter Reanalysis
Click Analysis Settingsthe Analysis Settings dialog box will pop up a. Adjust the start cycle and end cycle of the baseline b. Adjust Ct analysis algorithm c. Set up the use of S fitting d. Set up the stage to use for Ct analysis e. Set up the automatic threshold value f. Advanced setting g. Standard curve setting
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LineGene9600 Absolute Quantification
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LineGene9600 Absolute Quantification
5. Experiment Report
Start
This section describes how to print experiment report and covers report template designing and print setting.
Design experiment
Prepare for the reaction
Run the experiment
Experiment analysis
Experiment report
Data export
End
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LineGene9600 Absolute Quantification
5.1 Report Template Designing
Click Report Report Template Editorthe Report Designer window will pop up
The report consists of controls and the user can add, modify and delete controls.
Available controls include Static Text, Dynamic Text, Line, Static Image, Aamplification
Curve and Quantification Aanalysis Result.
5.2 Print Setting
Click Report Print Template Settingthe Print Template Setting window will pop up
The user can set up the hospital name, report name, reference value, tester, checker, amplification plot setup, default report template and paper size.
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LineGene9600
5.3 Report Printing
Click ReportReport Print
Report print setting a. Set up report template b. Print setting (please refer to Section 5.2) c. Select print items d. Print preview e. Print the report
Absolute Quantification
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LineGene9600 Absolute Quantification
6. Data Export
Start
This section describes how to export data and covers exporting to the database, experiment filing and exporting the experiment data to EXCEL
Design experiment
Prepare for the reaction
Run the experiment
Experiment analysis
Experiment report
Data export
End
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LineGene9600 Absolute Quantification
6.1 Export to Database
Click Data SummaryExport to Databasethe save file dialog box will pop upsave the exported database file
6.2 Experiment Filing
Click Data Summary Archived Experiment Diretory the Experimental archive storage directory window will pop upset up the storage path of file
Experiment filing
Click Data SummaryArchived Experiment export the filed experiment file
※
The suffix of filed experiment file is .fqh
6.3 Export Experiment Data to EXCEL
Click Data Summary Export Experiment to EXCEL set up Well Horizontal (set up
Well Vertically) the exported experiment data will generate EXCEL file
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LineGene9600
Chapter 5 Tool Usage
1. Gain Setting
Gain Setting tools are used to set up gain modes.
Click Tools Gain Settingthe following window will pop up
Gain setting can be set up as: reference gain, custom gain and auto gain
※
In the mode of Custom Gain, the user can modify the gain value.
Tool Usage
2. Target Library
Target Library tools are used to set up the inspection libraries of absolute quantitative.
Click Tools Target Library open the following window
The user can: a. Add Detector b. Modify Detector c. Delete Detector
3. Customized Dyes
Customized Dyes tools are used to set up existing dyes and newly added dyes.
Click Tools Customize Dyes open the following window
The user can: a. Create dye b. Modify dye name and channel c. Delete dye
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LineGene9600 Tool Usage d. Move dye upward e. Move dye downward
※
After adding new dyes or modifying dyes, the user should conduct crosstalk parameter measurement.
4. Customize Columns
Click Tools Customize Columns the following window will pop up
The user can: a. Add columns b. Delete columns c. Modify column name
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LineGene9600 Tool Usage
5. Column Selection
Select Columns tools are used to add the new columns in above section into current existing columns, or remove existing columns in current column.
Click Tools Select Columns the following window will pop up
※ a. Current existing column items include sample, absolute report, absolute report setting, query and query condition b. Double click column can add or remove a column c. Column with (*) indicates it cannot be removed
6. Sample Column Library
Sample Column Library tools are used in the experiment design phase. The user can selection the definition of contents in the drop-down box when setting up sample information.
Click Tools Sample Column Library the following window will pop up
The user can: a. Add columns b. Delete columns c. Edit the columns content
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LineGene9600 Tool Usage
7. Instrument Calibration Parameters
Instrument Calibration Parameters tools are used to calibrate instrument parameters.
Click Tools Instrument Calibration Parametersthe following window will pop up
8. Measure Crosstalk Calibration Parameters
Measure Crosstalk Calibration Parameters tools are used to measure crosstalk correction parameters.
Click Tools Measure Crosstalk Calibration Parameters the following window will pop up
The user can add and modify the channels to be tested and dyes according to his needs;
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LineGene9600 Tool Usage upload corresponding reaction plates and operate the experiment; when the experiment is over, the system will automatically save the crosstalk correction parameters.
9. Crosstalk Gain Parameter Measurement
Crosstalk Gain Parameter Measurement tools are used to measure crosstalk gain parameters.
Click ToolsMeasure Crosstalk Gain Parametersthe following window will pop up
The user can add and modify the channels to be tested and dyes according to his needs; upload corresponding reaction plates and operate the experiment; when the experiment is over, the system will automatically save the crosstalk gain parameters.
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LineGene9600 Tool Usage
10. System Maintenance
System Maintenance tools are used for system maintenance.
Click Tool Menu System Maintenancethe Password Input box will pop upinput the correct passwordsconduct the following setting: a.Y-axis commissioning b. X-axis origin calibration c. Machine serial number setting d. Photomultiplier setting e. Runtime zero clearing f. Background measurement g. Reference gain measurement h. Fluorescence incremental calibration
11. Firmware Upgrading
Firmware Upgrading tools are used to upgrade the firmware.
The instrument by connection of RS232 communication wire with computer realizes its software update as follows:
Set its rear MODE update switch of communication box on the back of the instrument to right side Update, turn on power supply and connect serial port line. The instrument is in update status, In the panel, indicator light and trouble light are lighted in the same time, which is normal.
Click Tool MenuFirmware Upgradingthe following window will pop up
The user can:
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LineGene9600 a. Select serial ports b. Select the BIN file to be upgraded c. Upgrade
Tool Usage
12. Convert Old File into New File
Convert Old File into New File tools are used to convert old files with the suffix of fqj or fqs into new files with the suffix of fqd.
Click Tool Menu Convert Old File into New Filethe following window will pop up
The user can: a. Add files to be upgraded b. Delete selected files c. Select the output directory of new files d. Upgrade
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LineGene9600 Other Functions
Chapter 6 Other Functions
1. Instrument Operation
Instrument operations include connecting the instrument, disconnecting the instrument and instrument information.
1.1 Connect Instrument
Click InstrumentConnectselect port number or select automatic port matching
When the instrument is connected, the icon on the status bar will be is disconnected, the icon on the status bar will be .
1.2 Disconnect
Click Instrument Disconnectdisconnect currently connected instrument
; if the instrument
1.3 Instrument Information
When the instrument is connected, the user can check the instrument information.
Click InstrumentInstrument Informationthe following dialog box will pop up
Instrument information includes instrument serial number, runtime, currently connected ports, whether the experiment is in operation.
2. Data Query
Data Query is used to query the data already exported to the database.
Click Data Summary Data Querythe following window will pop up
The user can: a. Select database files
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LineGene9600 b. Set up query condition c. Query d. Clear all query conditions
Other Functions
3. Open and Save File
Open a file
Click FileOpenthe Open File dialog box will pop up
The user can open experiment file (*.fqd), experiment template file (*.fqt), experiment filing file (*.fqh) and quantitative experiment file (*.fqj) in old version.
Save file
Click File(Save/Save As/Save As Template)
4. System Help
Click Help Help Topics
5. About
Click Help About the following window will pop up
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LineGene9600 Maintenance
Chapter 7 Maintenance
1. Regular cleaning
In order to ensure normal operation, detection and use, the instrument shall be regularly cleaned.
To clean of outer surface: It shall be cleaned with soft cloth, and if necessary, soft cloth may be soaked with alcohol, distilled water or cleaning paste.
To clean of module well: It may be cleaned with nail wipes which does not bring dust accompanied brush and clean soft cloth, and if necessary, they may be soaked with 95 percenta little of absolute ethyl alcohol used in medicine alcohol or distilled water.
Warning!
1. Before cleaning this instrument, the power supply shall be cut off.
2. At cleaning conical holes of the module, cares shall be take to prevent cleaning agent from dropping into the well.
3. The surface of this instrument shall not be cleaned with corrosive cleaning agent.
4. In order to avoid to scratch or damage optics in the wells, sharp and hard things are prevented to reach to wells of block.
Software update
The instrument by connection of RS232 communication wire with computer realizes its software update as follows:
Set its rear MODE update switch of communication box on the back of the instrument to right side Update, turn on power supply and connect serial port line. The instrument is in update status, In the panel, indicator light and trouble light are lighted in the same time, which is normal.
Open LineGene 9600 software, select absolute quantification function module, and from the main interface of absolute quantification module click “Firmware Upgrade (F)…” of “System
Parameters (P)” pull-down menu.
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LineGene9600 Maintenance
On the pop up dialog interface select the communicationserial port connecting instrument with computer.
Click “Load” and select update software (*.bin), which may be obtained from BIOER network or after service department.
To press “Begin” key to start update, if communication is successful, prompt sound will be heard.
and Aafter the update, “Update Completed” interface will pop up, prompt sound will be heard..
Press “Exit” key to complete software update..
After power off, sSet the update switch to Normal at left side and normally use instrument after power on.e to end the host software update.
2.
Analysis and elimination of troubles
No.
Trouble
1
2
3
The power switch at rear of instrument is set ON, but the instrument makes no response.
The module temperature runs normal, but the fluorescence detection makes no scanning
Display of system parameters menu requires input of
“Password”.
Possible cause
The run switch at front of instrument is not pressed, which shall be set “On” during running; otherwise the inside switching power will have no voltage output.
Correction
For working of instrument, its run switch shall be pressed and the green loop lamp shall light on to indicate energizing of the control system.
The fixing clamp is not removed or is not inserted and tightened to rear UNLOCKER well.
The system parameters are for instrument manufacturer’s inside calibration and require special accession password.
Remove fixing clamp and tighten to UNLOCKER well.
(For transport, the instrument shall normally shutdown and fixing clamp shall be removed and then screwed into side well.)
The function is unnecessary for the user and for calibration it shall contact manufacturer’s service personnel.
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LineGene9600 Maintenance
4
5
6
7
After turning on power switch the power lamp fails to light on
At detecting sample position, the step motor fails to work and the communication fails.
After detecting sample position, the actual temperature displays 0°C or
100°C
Module temperature/raising/lo wering rate obviously decreases or temperature control is incorrect.
The run switch is not pressed
Power is not switched on.
Burnt fuse
Damaged switch other
This switch is used for temporary turning on/off output power and is equivalent to “standby” function. For prolonged shutdown, the rear power switch shall be set “Off”.
Check and switch on power
Replace fuse (250V 8A
Ф5x20)
Replace switch
Contact the supplier or manufacturer
Check, connect or replace interface wire
Poor contact or damage of interface wire
The power switch is not turned on or is turned on only after program running
The step motor or the drive is damaged
The fixing clamp is not fully inserted.
Turn on power switch and restart program
Contact the supplier or manufacturer
Tighten it to UNLOCKER well, and re-turn on power after shutdown
The module temperature sensor is damaged. It accompanies panel red lamp alarm and software prompt, and instrument automatically stops running.
The power switch is turned on only after program running.
The program is searching communication port and during this period data would be not sent.
The ventilation opening is blocked.
Loose connection wire
The refrigerating sheet is damaged
Fan is damaged or fails to run
The temperature sensor is damaged
Contact the supplier or manufacturer
Turn on power switch and restart program
If the trouble still exists after researching, contact the supplier or manufacturer
Clear the stem of the ventilation opening
Contact the supplier or manufacturer
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LineGene9600 Maintenance
8
9
10
11
12
13
14
The module fails to heat and refrigerate.
Abnormal temperature or fluorescence curve: straight line or lose of partial data
Yellow lamp on panel lights on
The hot-lid is impossible to heat
Under no test tube state, the fluorescence value difference between wells increases or the background value is very high.
Reagent evaporation
Signal crosstalk among channels
Inside of instrument is damaged
The refrigerating sheet is damaged
Contact the supplier or manufacturer
During hot-lid heating-up
Waiting until hot-lid temperature comes to target value.
When stopping running, module temperature holds down 30°C automatically.
After virus-killing re-install application software.
The running program is infected by virus
Computer configuration fails to meet requirements or the setup of communication port is unreasonable.
The module is not fully pushed in and the optic coupler fails to detect the module
Thermal-sensitive fuse is damaged
Loose plug-pieces
Heating elements of hot-lid is damaged
Temperature sensor of hot-lid is damaged
Configurating as per require ments
In case of failure after re-pushing in, contact the supplier or manufacturer
Contact the supplier or manufacturer
The test tube well or hot-lid is contaminated, or baseline******.b96 background parameters are mis-used.
There is the matter with quality of test tube, whose sealing is not tight.
Dye signal among channels exists crosstalk objectively.
Eliminating contamination
Each instrument shall correspond to baseline96 document.
After perennial use, offset would occur in the optical elements. In this case, contact the manufacturer to re-calibrate the background value.
To select applicable consumptive material which has more tight capability.
You can measure by using
“Crosstalk Measurement”, and save parameters to modify.
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LineGene9600
15
Fluorescence detection value
-abnormal
Maintenance
Irradiation by external strong light
During running of program, the hot-lid is opened
The photo-electric system is damaged
Close external light source
Close hot-lid (detection result unreliable)
Contact the supplier or manufacturer
Caution:
During the warranty period, arbitrary opening amplification instrument casing and inspection shall not be allowed. In case of above troubles requiring opening amplification instrument casing for inspection, please timely contact the supplier or manufacturer.
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LineGene9600
Appendix: LineGene 9600 series wiring
1 2
+ -
4 5
1 2 3 4
1 2 3 4
1 2 3 4 5 6 7 8 9 10
1 2 3 4 5 6 7 8
Wiring Diagram
Inlet with fuse
POWER INPUT: 100-240V~ 50/60Hz 600W
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LineGene9600
DATA1=DATA2=8
(0.72deg,25,12500,0.0288deg)
RUN/STOP-CURRENT:SET=ORIGIN
FUC-DIPSW:1P=OFF,2P=3P=4P=ON
Wiring Diagram
FFC7# FFC8#
2 3
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1 2 3 4 5
LineGene9600 Wiring Diagram
B3900
TR TR
1 2
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Note:
Note:
Note:
Note:
Note:
Note:
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Questions & Answers
T T T
How does this device read fluorescence signals?
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Table of contents
- 10 CHAPTER 1 GENERAL DESCRIPTION
- 13 CHAPTER 2 PREPARATIONS
- 13 3.1 Removal of fixing pin
- 14 3.2 Connection of power wire and communication wire
- 14 4.1 System Installation
- 14 4.2 System Unloading
- 15 CHAPTER 3 START
- 17 CHAPTER 4 ABSOLUTE QUANTIFICATION
- 18 1.1 Create New Absolute Quantitative Experiment
- 18 Detector Setting
- 19 1.3 Sample Information Setting
- 20 1.4 Reaction Plate Setting
- 22 1.5 Program Setting
- 26 3.1 Preparation for reagent sample
- 26 3.2 Start Run
- 27 3.3 Monitor Operation
- 28 3.4 Program Setting
- 31 4.1 Check Results
- 36 4.2 Adjust Parameter Reanalysis
- 39 5.1 Report Template Designing
- 39 5.2 Print Setting