PikoReal User Manual_low res
Thermo Scientific PikoReal
Real-Time PCR System
User Manual
Rev. 2.0
sharing knowledge
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Thermo Scientific PikoReal
Real-Time PCR System
User Manual
Rev.2.0, Cat. no. N11471
Copyright
Copyright © 2011 Thermo Fisher Scientific Corporation. All rights reserved. Reproduction of the
accompanying user documentation in whole or in part is prohibited.
Licensing information
Not available in Canada, Brazil, UK, Germany, Austria, Switzerland, Italy, Spain, France, Belgium, the
Netherlands, Liechtenstein, Denmark and Sweden.
Notice: Purchase of this instrument conveys a limited, non-transferable immunity from suit for the purchaser's
own internal research and development and applied fields other than human in vitro diagnostics under one or
more of Canadian Patent 1,339,653, U.S. Patent 5,475,610 (claims 160-163 only) or corresponding claims in
their unexpired non-U.S. counterparts owned by Applied Biosystems. No right is conveyed expressly, by
implication or by estoppel under any other patent claim, including under U.S. Patent No. 6,814,934 or any
global equivalent thereof, which claims thermal cycler apparatus and systems capable of real-time detection.
Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied
Biosystems, 850 Lincoln Centre Drive, Foster City, California, 94404, USA.
Trademarks
“ALPS”, “DyNAmo”, “Finnpipette”, “Matrix”, “Maxima”, “Multidrop”, “Piko”, “PikoReal”, “Solaris”,
“Versette”, “Verso” and “UTW” are trademarks or registered trademarks of Thermo Fisher Scientific Inc. and
its subsidiaries.
All other trademarks and registered trademarks are the property of their respective holders.
Disclaimer
Thermo Fisher Scientific reserves the right to change its products and services at any time to incorporate
technological developments. This manual is subject to change without prior notice as part of a continuous
product development. Although this manual has been prepared with every precaution to ensure accuracy,
Thermo Fisher Scientific assumes no liability for any errors or omissions, nor for any damages resulting from
the application or use of this information. This manual supersedes all previous editions.
Remarks on screenshots
Screenshots may be slightly different on your system depending on the firmware version.
No liability for consequential damages
Thermo Fisher Scientific shall not be liable for any damages whatsoever arising out of the use or inability to
use this product.
Power failure
The system requires uninterrupted power supply in order to operate correctly. Thermo Fisher Scientific has no
responsibility whatsoever for system malfunctions arising from power failures.
About This User Manual
Intended users
About This User Manual
Intended users
The PikoReal Real-Time PCR System, as a standalone instrument or
with Thermo Scientific PikoReal Software, can be used in research
laboratories by professional personnel.
How to use this
user manual
This user manual is for the following instruments: 24-well PikoReal
Real-Time PCR System (Cat. no. TCR0024) and 96-well PikoReal
Real-Time PCR System (Cat. no. TCR0096). It aims to give you the
information you need for:
●
Reviewing safety precautions
●
Installing the PikoReal Real-Time PCR System
●
Operating the instrument
●
Adjusting the settings and servicing the instrument
●
Troubleshooting the instrument
This user manual also describes all the features and specifications of the
PikoReal Real-Time PCR System, and provides ordering information
for instruments, spare parts, reagents and consumables.
Read the manual in its entirety before operating the instrument.
Keep the user manual for future reference. The user manual is an
important part of the instrument and should be readily available.
For more
information
For PikoReal Software related issues, refer to the Thermo Scientific
PikoReal Software User Manual (Cat. no. N12076).
For the latest information on products and services, visit our website:
http://www.thermoscientific.com/pikoreal
In our efforts to provide useful and appropriate documentation, we
would appreciate any comments you may have on this user manual to
your local Thermo Fisher Scientific representative.
Thermo Fisher Scientific
Thermo Scientific PikoReal Real-Time PCR System User Manual
3
About This User Manual
Safety symbols and markings
Safety symbols
and markings
Safety symbols and
markings used on the
PikoReal Real-Time PCR
System
These symbols are intended to draw your attention to particularly
important information and alert you to the presence of hazards.
The following symbols and markings appear on the type label and the
instrument itself.
Power on ▲
Power off ▲
Serial number ▲
Catalog number ▲
Date of manufacture ▲
Consult instructions for use ▲
WEEE symbol This product is required to comply with the European
Union’s Waste Electrical & Electronic Equipment (WEEE) Directive
2002/96/EC. ▲
Two labels with the following texts (Figure 2–3):
ATTENTION!
This is shipping position only! The other hole is operating position,
refer to the manual. ▲
ATTENTION!
Always insert the plug into the Piko first, and then insert the power
cord into the wall outlet. ▲
4 Thermo Scientific PikoReal Real-Time PCR System User Manual
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About This User Manual
Safety instructions
Warning and other
markings used in
the documentation
The following symbols and markings appear in this user manual.
Warning Risk of electric shock. ▲
Warning Biohazard risk. ▲
Warning Hot surface, risk of burns. ▲
Warning Risk of injury to the user(s). ▲
Caution Risk of damage to the instrument, other equipment or loss of
performance or function in a specific application. ▲
Note Marks a hint, important information that is useful in the
optimum operation of the system, or an item of interest. ▲
Safety
instructions
Read the User Manual Operating the PikoReal Real-Time PCR
System without first reading the entire User Manual may constitute a
risk to your health. Only a person capable of handling electrical
equipment should use the PikoReal Real-Time PCR System. NOT
FOR CHILDREN. ▲
Do Not Attempt to Repair the System Do not remove the cover of the
system. Do not try to repair or replace broken components – you put
yourself at risk for electrical shock. Removing the cover or
replacing/removing components will void the warranty. Contact
customer service or your local distributor if your system is not
functioning properly. ▲
Do Not Touch the Sample Block Certain components, including the
sample block and heated lid, will become excessively hot. Touching
these components may cause burns. ▲
Do Not Tamper with Electronics Coming into contact with the
electronics, even when the system is off or unplugged, may cause an
electrical shock or harm. ▲
Do Not Use Flammable, Hazardous or Radioactive Liquids with the
PikoReal Real-Time PCR System. ▲
Thermo Fisher Scientific
Thermo Scientific PikoReal Real-Time PCR System User Manual
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About This User Manual
Safety instructions
6 Thermo Scientific PikoReal Real-Time PCR System User Manual
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Contents
Contents
Intended users ................................................................................... 3
How to use this user manual ............................................................. 3
For more information ....................................................................... 3
Safety symbols and markings............................................................. 4
Safety symbols and markings used on the PikoReal Real-Time PCR
System............................................................................................ 4
Warning and other markings used in the documentation ............... 5
Safety instructions ............................................................................. 5
Thermo Fisher Scientific
Chapter 1
Introduction to the PikoReal® Real-Time PCR System ........................ 11
Introduction to qPCR..................................................................... 13
Intended use.................................................................................... 13
Chapter 2
General Description .................................................................................... 15
Instrument views............................................................................. 15
PikoReal user interface .................................................................... 17
Status indicator ............................................................................ 17
Power supply................................................................................... 17
PikoReal Software ........................................................................... 18
Block formats .................................................................................. 18
Consumables................................................................................... 18
UTW technology ......................................................................... 19
Reagents.......................................................................................... 20
Dye calibration................................................................................ 21
Dispensers....................................................................................... 21
Heat sealers ..................................................................................... 21
Chapter 3
Installation .................................................................................................... 23
What to do upon delivery ............................................................... 23
Unpacking the instrument ........................................................... 23
Checking delivery for completeness and damage .......................... 23
Environmental requirements ........................................................ 24
Air flow ..................................................................................... 25
Installation steps.............................................................................. 25
Connecting the power supply cable .............................................. 25
Connecting to the network .......................................................... 26
Connecting to the data port ......................................................... 26
Warnings and cautions.................................................................... 26
Electrical ...................................................................................... 26
Operational check ........................................................................... 27
Installing PikoReal Software............................................................ 27
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Contents
Chapter 4
Routine Operation ........................................................................................29
Switching on ................................................................................... 29
Status indicator ............................................................................... 29
Main menu ..................................................................................... 29
Running a protocol ......................................................................... 30
Monitoring a run ............................................................................ 32
Status screen................................................................................. 32
Time screen.................................................................................. 33
Functions in the screens ............................................................... 33
Tools menu..................................................................................... 34
System info .................................................................................. 34
Diagnostics .................................................................................. 35
Lid ............................................................................................ 35
Sensors ...................................................................................... 36
Display...................................................................................... 36
Clock ........................................................................................ 37
Network....................................................................................... 37
Service.......................................................................................... 38
Logfiles......................................................................................... 38
Park instrument ........................................................................... 38
Run data to USB.......................................................................... 39
Shutting down ................................................................................ 40
Chapter 5
Maintenance .................................................................................................41
Regular and preventive maintenance ............................................... 41
Maintenance checklist .................................................................. 41
Instrument care ............................................................................ 41
Cleaning of the instrument ............................................................. 42
Cleaning the block and wells ........................................................ 43
Cleaning the heat sink.................................................................. 43
Disposal of materials ....................................................................... 44
Decontamination procedure............................................................ 44
Packing for service........................................................................... 45
Maintaining a system log................................................................. 46
Disposal of the instrument .............................................................. 46
Chapter 6
Technical Specifications ...........................................................................47
Instrument specifications................................................................. 47
Safety specifications......................................................................... 48
In conformity with the requirements............................................ 48
Chapter 7
Troubleshooting Guide................................................................................51
Handling abnormal situations ......................................................... 51
Power failure ................................................................................... 51
Warning and error codes ................................................................. 51
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Contents
Thermo Fisher Scientific
Chapter 8
Ordering Information................................................................................... 55
PikoReal.......................................................................................... 55
List of spare parts and accessories .................................................... 55
List of consumables ......................................................................... 56
List of reagents ................................................................................ 57
List of heat sealers ........................................................................... 57
Appendix A
System Log .................................................................................................... 59
Appendix B
Certificate of Decontamination................................................................. 61
Thermo Scientific PikoReal Real-Time PCR System User Manual
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Contents
10 Thermo Scientific PikoReal Real-Time PCR System User Manual
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Introduction to the PikoReal( Real-Time PCR System
Safety instructions
Chapter 1
Introduction to the PikoReal®
Real-Time PCR System
The PikoReal Real-Time PCR System (Figure 1-1) is a gene
quantification and genotyping platform in 24- and 96-well block
formats. Using PikoReal Software, the system is applicable to absolute
quantification, relative quantification, melt curve analysis and allelic
discrimination. The PikoReal Real-Time PCR System is designed to a
minimal footprint, making the system ideal for personal benchtop use
and field applications, for example.
Figure 1-1. PikoReal 96 Real-Time PCR System
The combination of several innovative technologies applied in design,
including optics, unique heat sink design, block design, plate size and
the usage of special plasticware, create many advantages over
conventional qPCR systems. The PikoReal Real-Time PCR System is
fast, fulfills high-level requirements of temperature uniformity over the
wells and creates significant savings in reagent, plastic material and
energy consumption.
The PikoReal Real-Time PCR System uses ultra-thin wall (UTW®)
microwell plates that are 1/4 of the size of conventional microwell
plates. The plates adhere to industry standards regarding well spacing
and sample capacity and are compatible with multichannel pipettes,
Thermo Fisher Scientific
Thermo Scientific PikoReal Real-Time PCR System User Manual
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Introduction to the PikoReal( Real-Time PCR System
Safety instructions
reagent dispensers and automated liquid handling systems. The
PikoReal 24 also utilizes standard low profile 8-well strips and 0.2 ml
tubes.
The optical system comprises a charge-coupled device (CCD) camera,
optical filters and an individual light-emitting diode (LED) for each of
the five channels. The channel selection is built on a motorized linear
shuttle that moves to five channel positions.
The PikoReal is controlled by PikoReal Software via an Ethernet
connection (with up to 10 connections) or alternatively as a standalone
instrument with USB memory stick loaded protocols.
The PikoReal Real-Time PCR System has the following features:
●
Compact design – The PikoReal Real-Time PCR System has a
compact design giving a minimal footprint for personal benchtop
use and field applications.
●
Unique thermal system – Three Peltier elements and a patented
heat sink design provide a fast and even heat distribution and
removal.
●
5 LED optical system – Separate LEDs for each channel offer a
wide excitation range (475–640 nm) and multiplexing for four
dyes.
●
Piko® format – The PikoReal Real-Time PCR System sample
blocks (24-well and 96-well) will only accept microplates with a
footprint with the dimensions of a microscope slide – the Piko
format. The 24-well sample block is also compatible with all
standard low-profile 8-tube strips and single tubes.
●
Automatic lid – The heated lid is internally motor-controlled and
the lid will automatically adjust to the optimal temperature and
sealing pressure.
●
External power supply – The PikoReal Real-Time PCR System
uses an external 200 watt power supply (an approach commonly
used with lap-top computers).
Note Conventional format full-sized microplates are not compatible
with the PikoReal blocks. ▲
The PikoReal Real-Time PCR System is available in two different
configurations:
●
●
PikoReal 24
•
Cat. no. TCR0024
•
for use with 24-well Piko PCR plates, standard 8-well strips
and 0.2 ml tubes
PikoReal 96
•
12 Thermo Scientific PikoReal Real-Time PCR System User Manual
Cat. no. TCR0096
Thermo Fisher Scientific
Introduction to the PikoReal( Real-Time PCR System
Introduction to qPCR
•
for use with 96-well Piko PCR plates.
The following components form the complete technology platform for
performing real-time PCR based DNA analyses:
Introduction to
qPCR
●
PikoReal Real-Time PCR System
●
PikoReal Software (see “PikoReal Software” on page 18)
●
UTW® consumables (see “Consumables” on page 18)
●
qPCR reagents (see “Reagents” on page 20).
Quantitative real-time PCR (qPCR) is a method for analyzing DNA by
detecting the amount of PCR product after each PCR cycle by the
incorporation of fluorescent dyes into the reaction. The amplification
and detection of a specific DNA sequence is integrated into a
simultaneous process within a single instrument. The product
accumulation is detected and measured throughout the reaction – in
real-time. The detection is based on monitoring fluorescent molecules
that are incorporated into the amplified products as a dsDNA binding
dye or within labeled target specific primers or probes.
The quantitation is based on correlation between the measured
fluorescence intensity and the amount of DNA formed. Collecting
quantitative data of a known dilution series of the target DNA can be
used to give insight into the PCR reaction kinetics and reaction
efficiency. The gene quantification analysis method can be absolute
(copy number, concentration) or relative (to a known reference gene
and calibrator sample). Examples of qualitative analysis include gene
identification and genotyping, e.g. studying DNA sequence variations,
such as single nucleotide polymorphism.
The basic difference from end-point PCR technique is that in addition
to the qualitative result – detecting the presence of a PCR product, a
quantitative result – the initial amount of the amplified product – can
be determined. qPCR offers many advantages over traditional endpoint PCR methods: high sensitivity over a broad dynamic range,
reproducibility, low contamination risk and the exclusion of post-PCR
manipulations for analysis. It also facilitates high-throughput and
multiplexing applications.
Intended use
The PikoReal Real-Time PCR System is a thermal cycler with a
fluorescence measuring system available in 24-well and 96-well formats.
It is intended for qPCR technologies and related fluorogenic
chemistries. The PikoReal Real-Time PCR System is intended for
research purposes only.
Use for any other than the above-mentioned purposes is excluded.
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Introduction to the PikoReal( Real-Time PCR System
Intended use
Caution Operate the instrument only with software and hardware that
is specifically designed or selected for it. Thermo Fisher Scientific
assumes no liability for the use of third-party software applications. ▲
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General Description
Instrument views
Chapter 2
General Description
This chapter describes the main parts of the PikoReal instrument.
Instrument
views
The front view of the PikoReal instrument is shown in Figure 2–2.
User interface
Data port
Air exhaust
Drawer
Air intake
Feet
Figure 2–2. PikoReal front view
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General Description
Instrument views
The rear view of the PikoReal instrument is shown in Figure 2–3.
Air exhaust
Type label
Power
connector
Ethernet port (RJ45)
Figure 2–3. PikoReal rear view
Caution Do not block the air intake or the air exhaust vents
(Figure 2–2 and Figure 2–3). Be careful not to place, for example, a
piece of paper under the air intake. ▲
Warning Do not place anything in front of the drawer. Items may be
knocked over when opening the drawer. ▲
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General Description
PikoReal user interface
PikoReal user
interface
The PikoReal user interface (UI) and its functions are shown in
Figure 2–4.
Status indicator: Blue
Red
Power button
Backlit LCD display
Arrow keys for
moving on
the display
Cancel key
Enter key
Open/Close
drawer button
Figure 2–4. PikoReal user interface
Status indicator
On the top left corner of the instrument user interface is a status
indicator light that has the following states (Table 2–1):
Table 2–1. Status indicator
Upper light
Lower light
Power supply
Indicator
Status
Constant blue
Instrument is on but not in use.
Blinking blue
Instrument is on and in use.
Constant red
System error. For more information, see Chapter 7:
“Troubleshooting Guide”.
The PikoReal Real-Time PCR System is equipped with an external
200 watt power supply.
A green light on the power supply indicates normal operation.
The power supply will get warm during normal operation.
Note If the internal circuit breaker trips, the green light of the power
supply will go off. The power supply must be unplugged from the
power outlet for 5 minutes to reset it. ▲
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General Description
PikoReal Software
Note Do not cover the power supply with anything as this will impede
the ability of the power supply to efficiently dissipate heat. ▲
PikoReal
Software
Block formats
The PikoReal is operated with the PikoReal Software application. Up to
ten PikoReals can be simultaneously operated from a single computer.
For more information, refer to the PikoReal Software User Manual
(Cat. no. N12076).
The PikoReal Real-Time PCR instrument is available in two block
formats (Figure 2–5 and Figure 2–6).
Figure 2–5. 24-well, 50 µl max. volume for PCR
Figure 2–6. 96-well, 20 µl max. volume for PCR
Consumables
PikoReal 24-well and 96-well formats utilize Piko PCR plates, which
are 1/4th the size of standard PCR plates. See Figure 2–7.
The plates maintain the industry standard for well volumes and well
spacing, ensuring the compatibility with standard multi-channel
pipettes (both 8- and 16-channel) and liquid handling robots.
Four 24-well plates are equivalent to a standard ANSI 96-well plate and
four 96-well Piko PCR plates are equivalent to a standard ANSI 384well plate.
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General Description
Consumables
Plate frames are available to assemble Piko PCR plates into standard
ANSI format plate size (see Chapter 8: “Ordering Information”).
The 24-well Piko PCR plates are sealed with strip caps or with a heat
sealing film, whereas 96-well Piko PCR plates are sealed with a heat
sealing film.
The 24-well sample block is also compatible with low profile 8-tube
strips (sealed with attached* or separate cap strips) and single tubes with
flat optical caps.
* Not available in US.
Note If you are using single tubes, you must use at least eight tubes that
are placed symmetrically with at least one tube in each corner of the
block.
You can use empty tubes to fill the above requirement when analyzing
fewer samples. ▲
Figure 2–7. Piko PCR plates
UTW technology
Thermo Fisher Scientific Piko PCR plates, strips and tubes utilize
UTW (ultra-thin wall) technology and are recommended to be used
with PikoReal.
UTW reaction vessels were used in the optimization of the temperature
algorithms of the instrument as well as in the validation of its
performance.
Note For the highest signal-to-background ratio and elimination of
possible well-to-well cross-talk, white plates, strips and tubes are
recommended. ▲
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General Description
Reagents
The UTW consumables are half the thickness (< 0.15mm) of
conventional PCR consumables and thus improve the thermal transfer
between block and sample (see Figure 2–8 and Figure 2–9).
The uniform tube design allows repeatable and consistent results from
well to well.
Figure 2–8. The tube wall thickness of UTW consumables (marked as blue)
is half that of standard consumables (marked as red).
UTW area
Figure 2–9. The lower portion of the UTW reaction vessels is ultra-thin.
For more information on the consumables, refer to Chapter 8:
“Ordering Information” and http://www.thermoscientific.com/pikoreal.
Reagents
Thermo Scientific offers a comprehensive selection of qPCR reagents to
be used with the PikoReal Real-Time PCR System. These include, for
example, universal reagents for SYBR Green and probe chemistries,
SNP Genotyping as well as Solaris pre-designed qPCR assays for geneexpression analysis.
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General Description
Dye calibration
For more information on the reagents, refer to Chapter 8: “Ordering
Information” and http://www.thermoscientific.com/pikoreal.
Dye calibration
The PikoReal has been precalibrated to use the following fluorophores:
●
FAMTM
●
HEXTM
●
Yakima Yellow®
●
ROXTM
●
Texas Red®
●
Cy®5
The PikoReal Real-Time PCR System is also compatible with SYBR®
Green I and similar DNA intercalators. If you wish to calibrate other
dyes into the system, refer to Appendix A: “Color Calibration” in the
PikoReal Software User Manual (Cat. no. N12076).
Dispensers
Thermo Fisher Scientific provides a variety of dispensers compatible
with Piko PCR plates: Finnpipette® and Matrix® multichannel pipettes,
adjustable electronic Matrix® pipettes as well as Multidrop® Combi
reagent dispensers and Versette™ automatic liquid handlers.
For more information on Thermo products, refer to
http://www.thermoscientific.com.
Heat sealers
Thermo Fisher Scientific provides a variety of ALPS ™ heat sealers to be
used with heat sealing films.
For more information on heat sealer products, refer to Chapter 8:
“Ordering Information” or http://www.thermoscientific.com and
http://www.matrixtechcorp.com.
Thermo Fisher Scientific
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General Description
Heat sealers
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Installation
What to do upon delivery
Chapter 3
Installation
This chapter describes the installation of the PikoReal Real-Time PCR
System.
What to do upon
delivery
Unpacking the
instrument
This section covers the relevant procedures to be carried out on receipt
of the instrument.
Move the packed instrument to its site of operation. To prevent
condensation, the instrument should be left in its protective plastic
wrapping until the ambient temperature has been reached.
Note Unpack the PikoReal Real-Time PCR System according to the
separate instructions included in the package. ▲
Unpack the PikoReal Real-Time PCR System and its accessories
carefully so that the arrows on the transport packaging point upwards.
Place the instrument onto a laboratory bench.
Caution Do not touch or loosen any screws or parts other than those
specifically designated in the instructions. Doing so may cause
misalignment and will void the instrument warranty. ▲
Retain the original packaging for future transportation or in case you
need to return the PikoReal Real-Time PCR System for maintenance.
The packaging is designed to ensure safe transport and minimize transit
damage. Use of insufficient packaging materials may invalidate the
warranty. Retain all instrument-related documentation provided by the
manufacturer for future use.
If you relocate your instrument or ship it for service, refer to “Packing
for service” on page 45.
Checking delivery
for completeness
and damage
Thermo Fisher Scientific
Check the enclosed packing list against the order. Check that the
delivery includes the following items:
●
PikoReal Real-Time PCR System
●
PikoReal Software for PikoReal Real-Time PCR System CD
Thermo Scientific PikoReal Real-Time PCR System User Manual
23
Installation
What to do upon delivery
●
Power supply
●
Power cord
●
Ethernet cable
●
Unpacking/packing instructions
Note that a personal computer is not provided by Thermo Fisher
Scientific. A PC is required for full use of the PikoReal functionalities.
For recommended PC requirements, refer to the PikoReal Software User
Manual (Cat. no. N12076).
Visually inspect the transport package, the instrument and the
accessories for any possible damage. If any parts are missing or damaged,
contact your local Thermo Fisher Scientific representative or Thermo
Fisher Scientific Oy.
Environmental
requirements
When setting up your PikoReal Real-Time PCR System, avoid locations
with excessive dust, vibrations, strong magnetic fields, direct sunlight,
draft, excessive moisture or large temperature fluctuations.
The instrument will perform optimally and exhibit the highest reliability
within the recommended environmental ranges (Table 3–2). Although
the instrument is rated to operate reliably within the absolute
environmental ranges, avoid the extremes to best preserve the long-term
performance and life span of the instrument.
Table 3–2. Operating conditions
Environmental condition
Absolute ranges
Recommended ranges
Ambient temperature
5–30°C
15–25°C
Ambient relative humidity
below 80%
below 50%
Altitude
below 2000 m
below 1000 m
Make sure that:
●
The working area is flat, dry, clean and vibration-proof, and leaves
additional room for cables, covers, etc.
●
There is sufficiently room behind the instrument to enable
disconnecting the device.
●
The ambient air is clean and free of corrosive vapors, smoke and
dust.
●
The ambient temperature range is between +5°C and +30°C.
●
The humidity is low so that condensation does not occur (relative
humidity is between 10% and 80%).
Caution Do not operate the instrument in an environment where
potentially damaging liquids or gases are present. ▲
24 Thermo Scientific PikoReal Real-Time PCR System User Manual
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Installation
Installation steps
Note To ensure that your PikoReal Real-Time PCR System will
provide years of top-level performance and will have the fewest
problems, adhere to the given environmental requirements. ▲
Air flow
Airflow is paramount to high-speed performance and high reliability.
Maintain good heat sink cooling capability by ensuring a cool source of
incoming air and an unobstructed exhaust of air.
To ensure obstruction-free intake of air:
●
Do not set the instrument on a hot surface or on laboratory bench
paper.
●
Do not slide any paper or other material under the instrument as
this may hamper the airflow or be sucked into the instrument.
To ensure obstruction free exhaust of air:
●
Always keep at least 10 cm of distance between the exhaust vents
and any large solid objects or walls.
●
Do not have other instruments exhaust directly at the PikoReal
Real-Time PCR System.
Clean the fins of the heat sink and the instrument regularly (see
“Cleaning the heat sink” on page 43).
Installation
steps
This section describes the installation steps that have to be carried out
before operating the instrument.
Caution If the instrument was stored or transported in a cold
environment, allow it to reach room temperature before plugging it in.
Leave the instrument to sit for at least three hours before installing and
switching it on, to prevent condensation causing a short circuit. ▲
Connecting the
power supply cable
To connect the power supply cable:
Warning Never operate your instrument from a power outlet that has
no ground connection. Never use a power supply cable other than the
Thermo Scientific power supply cable designed for your region. ▲
1. Plug the power supply into the back of the PikoReal Real-Time
PCR System (Figure 3–10).
Caution To prevent risk of shock or damage to the instrument, always
connect the power supply first to the instrument before plugging it into
the power outlet. ▲
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Installation
Warnings and cautions
2. Insert the power cord into the power supply.
3. Insert the power cord into the power outlet.
The system will power up automatically.
Power connector
Ethernet port (RJ45)
Figure 3–10. Connecting the power supply cable and the Ethernet cable
Connecting to the
network
Connect the Ethernet cable to the back of the PikoReal Real-Time PCR
System (Figure 3–10). You must use a shielded Ethernet cable
(Cat5e/Cat6e, FTP/STP). For more information, refer to the PikoReal
Software User Manual (Cat. no. N12076).
Connecting to the
data port
The data port (see Figure 2–2 on page 15) is to be used with USB
memory devices only. For instructions on how to use a USB memory
device, refer to “Running a protocol” on page 30 and “Run data to
USB” on page 39.
Warnings and
cautions
This instrument is designed to be safe for use. When correctly installed,
operated and maintained, it will present no hazard to the user.
The following recommendations are given for added user safety.
Electrical
Ensure that the power supply cable supplied with the unit is always
used. If a correct type of mains cable is not provided, use only cables
certified by the local authorities.
26 Thermo Scientific PikoReal Real-Time PCR System User Manual
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Installation
Operational check
The power plug should only be inserted into a socket outlet with a
protective ground contact. Never use an extension cable without a
protective ground wire. Always ensure that the electricity supply in the
laboratory conforms to that specified on the type label of the
instrument.
Warning Only authorized technical service personnel are allowed to
open the instrument. ▲
Warning Do not touch switches or electrical outlets with wet hands.
Switch the instrument off before disconnecting it from the mains
supply. ▲
Operational
check
Before you put the instrument into use, perform the following
operational check:
1. Plug the instrument into the mains. The instrument automatically
proceeds to perform the self tests. The self tests are performed each
time the instrument is turned on.
2. After approximately 30 seconds a beep signal indicates the test
completion.
3. The operational check has passed if no error messages appear. In
case of errors, refer to “Warning and error codes” on page 51.
Note Partial self tests are performed every time a protocol is started. ▲
Installing
PikoReal
Software
Thermo Fisher Scientific
The PikoReal Software is on the Thermo Scientific PikoReal Real-Time
PCR System CD (Cat. no. N12119). The CD also contains the
instrument and software user manuals.
For installation instructions, refer to the PikoReal Software User Manual
(Cat. no. N12076).
Thermo Scientific PikoReal Real-Time PCR System User Manual
27
Installation
Installing PikoReal Software
28 Thermo Scientific PikoReal Real-Time PCR System User Manual
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Routine Operation
Switching on
Chapter 4
Routine Operation
The operation of the PikoReal Real-Time PCR System is described
below.
Switching on
Before switching the instrument on, check that all the cables are
properly fitted according to the installation instructions. Note that when
you plug the instrument into the mains, it automatically starts.
If the instrument is plugged in and on standby mode, switch the
instrument on by pressing the POWER button.
After startup, the instrument lid pre-heats automatically. After three
hours’ inactivity, the lid pre-heating is turned off.
Status indicator
Upper light
Lower light
Main menu
To the left side of the power button on the keypad are two indicator
lights with the following indications:
Constant blue
System is on and NOT in use.
Blinking blue
System is on and in use.
Constant red
System has recently experienced an error or warning. Refer to
Chapter 7: “Troubleshooting Guide” for more information.
The PikoReal user interface (see Figure 2–4) features four function keys
which are used to select different menu options.
The function fields on the Main menu have the following actions:
RUN
Runs a protocol from USB or instrument memory.
STAT
Shows the Status screen (when running).
TOOLS
Accesses global settings or diagnostic routines.
[C] The red button with a “C” is the CANCEL button. It will either bring
you back to the previous menu or it will cancel your run immediately
when you continuously hold down the button. Pressing “C” also clears
a message from the display.
Thermo Fisher Scientific
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Routine Operation
Running a protocol
↵ Selects the highlighted item.
Running a
protocol
The protocols can be run using PikoReal Software (and Ethernet
connection) or from a USB memory stick. The last protocol, which was
run using the software, can also be rerun from the instrument memory.
To run a protocol using PikoReal Software, refer to the PikoReal User
Software Manual (Cat. No. N12076). To run a protocol from USB or
from the instrument memory:
1. Select RUN in the Main menu and press Enter.
2. If a USB is attached, the USB folder will appear and is highlighted
as default folder. The last performed protocol , which was run using
the software, is stored in the [Recent] folder (and also in the
[Shared] folder).
3. Select a folder and press Enter. The protocols stored in that folder
will be displayed.
4. Select the protocol and press Enter. The protocol will be displayed
in semi-graphical mode.
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Routine Operation
Running a protocol
Note Protocol files on the USB memory stick can have a name that is
up to eight characters long. ▲
Note Thermo Fisher Scientific does not guarantee that all USB memory
devices are compatible with the instrument. ▲
5. The protocol can be viewed in list mode or graph mode, and
browsed using the Left/Right or the Up/Down arrow keys,
correspondingly.
6. After selecting the view mode (list or graph), start a run by selecting
START and pressing Enter.
With a 24-well PikoReal Real-Time PCR System, the users will be
presented with consumable and volume options. With a 96-well
PikoReal Real-Time PCR System, the users will only be presented with
volume options.
7. Select the consumable (that is, the vessel type) for a 24-well
PikoReal Real-Time PCR System. Select either:
●
< 16 TUBES if using 8–16 individual 0.2 ml tubes or
●
> 16 TUBES OR PLATE if using a 24-well Piko PCR Plate.
Caution To avoid risk of contaminating the instrument, always use
high-quality plastic consumables, such as UTW consumables. For more
information, refer to Chapter 8: “Ordering Information”. ▲
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Routine Operation
Monitoring a run
Note If less than 16 x 0.2 ml tubes have been loaded and > 16 TUBES
OR PLATE is selected by mistake, the lid pressure may cause the tubes
to become crushed or deformed. ▲
Note If you are using single tubes, you must use at least eight tubes that
are placed symmetrically with at least one tube in each corner of the
block. You can use empty tubes to fill the 8-tube requirement when
analyzing fewer samples. ▲
8. Enter the sample volume (µl per well) using the arrow keys.
●
24-well PikoReal: 10–50 µl
●
96-well PikoReal: 5–20 µl
9. Press Enter to confirm the desired volume.
Note When a run begins, the lid inside the instrument will begin
moving, a blue indicator light will flash, and the running protocol will
be displayed in semi-graphical mode on a Status screen (see
“Monitoring a run” on page 32). ▲
Monitoring a run
Status screen
You can monitor a run in two screens: Status screen and Time status
screen.
Upon initiation of a run, initialization tests are carried out and the
Status screen is displayed. The current temperature step in the protocol
is represented by a flashing line (graph view) or highlighted (list view),
and the protocol and host folder are displayed on the top of the screen.
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Routine Operation
Monitoring a run
Note If you press the C key during the run, the Main menu screen is
displayed with a STAT option. Selecting STAT returns the display to the
Status screen. ▲
Time screen
To enter the Time screen, select TIME and press Enter.
The digitals in the middle of the screen display the time (hours, minutes
and seconds) remaining until the completion of the running protocol.
The file name and host folders are displayed on the top of the screen.
The following status parameters are displayed:
Functions in the
screens
Thermo Fisher Scientific
●
FILE – Displays the path and filename of the current protocol.
●
STEP – Displays the step number, temperature and time of the
currently running temperature step.
●
CYCLE – Displays the current cycle number (within a GOTO
loop).
In both the Status and Time status screens, the following actions can be
used:
Thermo Scientific PikoReal Real-Time PCR System User Manual
33
Routine Operation
Tools menu
PAUSE
Pauses the run at the present temperature step, or at the next step if
ramping. When paused, this option becomes RESUME.
NEXT
Skips to the next protocol step.
TIME
Toggles to the Status screen.
STOP
Stops the protocol.
You can also return to the Main menu while the instrument is running
by pressing the C key on the keypad.
Tools menu
Select TOOLS in the Main menu, and press Enter.
The TOOLS menu provides six options, which are accessed using the
arrow keys and then by pressing Enter:
System info
●
SYSTEM INFO – Displays system info.
●
DIAGNOSTICS – Allows users to check lid parameters, sensor
parameters, display settings, and clock settings.
●
NETWORK – Allows users to view network address values for
networking PikoReal.
●
SERVICE – Used by authorized service personnel only!
●
LOGFILES – Displays protocol and instrument log files.
●
PARK INSTRUMENT – Prepares the instrument for
transportation.
●
RUN DATA TO USB – Appears when a USB memory stick is
inserted. Copies the data of the previous protocol to the memory
stick once the protocol is ready.
Select SYSTEM INFO in the TOOLS menu with the arrow keys and
press Enter.
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Routine Operation
Tools menu
The System Info screen displays information about the device and
software:
Diagnostics
●
TYPE – Displays the type of PikoReal instrument.
●
S/N – Displays the serial number of the device.
●
PIKO – Displays the firmware version of the PIKO module (i.e. the
thermal cycler module).
●
RT – Displays the firmware version of the RT module (i.e. the realtime module).
Select DIAGNOSTICS in the TOOLS menu with the arrow keys and
press Enter.
The Diagnostics screen displays data about the four following settings:
Lid
●
LID (see “Lid” on page 35)
●
SENSORS (see “Sensors” on page 36)
●
DISPLAY (see “Display” on page 36)
●
CLOCK (see “Clock” on page 37)
Select DIAGNOSTICS in the TOOLS menu with the arrow keys and
press Enter.
Select LID with the arrow keys and press Enter.
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Routine Operation
Tools menu
The Lid screen displays the offset settings for ZERO, LOW, MED
(=MEDIUM) and HIGH lid pressure. Users may be asked to read these
values to the service personnel as part of a troubleshooting procedure.
The user can drive the LID UP and DOWN by pressing Enter when
the offset is selected. Note that this is not possible during a run.
Sensors
Select DIAGNOSTICS in the TOOLS menu with the arrow keys and
press Enter.
Select SENSORS with the arrow keys and press Enter.
The Sensors screen displays sensor data from the device:
Display
●
BLOCK – Displays the current block temperature.
●
LID – Displays the current lid temperature.
●
SINK – Displays the current sink temperature.
●
DRAWER – Displays the state of the drawer (OPEN/CLOSED).
●
LID – Displays the state of the lid (UP/DOWN).
Select DIAGNOSTICS in the TOOLS menu with the arrow keys and
press Enter.
Select DISPLAY with the arrow keys and press Enter.
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Routine Operation
Tools menu
The display screen allows you to adjust the contrast of the PikoReal’s
LCD screen. The contrast is lowered or raised by pressing the Up and
Down arrow keys. The change in contrast will appear immediately.
Press Enter to accept the new contrast setting.
Clock
Select DIAGNOSTICS in the TOOLS menu with the arrow keys and
press Enter.
Select CLOCK with the arrow keys and press Enter.
The Clock screen allows viewing of the PikoReal’s internal date and
time settings.
Use the PikoReal Software (PC software) to modify the date and time.
Note If the PikoReal instrument is unplugged and without power for
more than 72 hours, you may have to reset the clock. ▲
Network
Select NETWORK in the TOOLS menu with the arrow keys and press
Enter.
The Network screen allows the user to view the IP address, the subnet
mask (MASK), the gateway (GW), the Domain Name Server (DNS),
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37
Routine Operation
Tools menu
and the MAC address (MAC). It is not possible to edit the MAC
address.
Note It is not possible to change the MASK, GW or DNS values
without help from authorized technical service. ▲
Use the PikoReal Software (PC software) to change the IP address of the
instrument, if necessary.
Service
Note that this function is to be used by authorized Thermo Fisher
Scientific service personnel only.
Logfiles
Select LOGFILES in the TOOLS menu with the arrow keys and press
Enter.
The Logfiles screen allows the user to view protocol and instrument
related log files:
Park instrument
●
PROTOCOL LOGS – Displays the log files of the last 10 executed
protocols.
●
INSTRUMENT LOG – Displays the instrument log files.
With the park instrument option, the user can prepare the device for
transportation.
Select PARK INSTRUMENT in the TOOLS menu with the arrow
keys and press Enter. Follow the instructions on the screen.
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Routine Operation
Tools menu
Note In the new models of the instrument the locking screw is not
required. This is indicated on the back panel, refer to Figure 2–3 on
page 16. ▲
When you restart the instrument, a prompt appears to check that you
have removed the transport locking screw from the shipping position.
Note In the new models of the instrument the locking screw is not
required. This is indicated on the back panel, refer to Figure 2–3 on
page 16. ▲
Run data to USB
Thermo Fisher Scientific
Select RUN DATA TO USB in the TOOLS menu with the arrow keys
and press Enter. Note that he USB memory stick has to be inserted.
Thermo Scientific PikoReal Real-Time PCR System User Manual
39
Routine Operation
Shutting down
Answer the prompt using the arrow keys and press Enter. This option
enables copying the backup data of the last performed protocol to the
memory stick once the protocol is ready. This option is disabled while
the protocol is active.
Note Thermo Fisher Scientific does not guarantee that all USB memory
devices are compatible with the instrument. ▲
Shutting down
To shut down the PikoReal Real-Time PCR System:
1. Remove any UTW vessels, tube strips or single tubes from the
instrument.
2. Answer the prompt using the arrow keys and press off the
instrument after use by holding down the POWER button
continuously.
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Maintenance
Regular and preventive maintenance
Chapter 5
Maintenance
Regular and
preventive
maintenance
Maintenance
checklist
Contact local authorized technical service or your local Thermo Fisher
Scientific representative for assistance, if necessary.
This chapter contains an outline of the points mentioned in the
checklist below (Table 5–3). Routine and service procedures must be
performed by the user to prevent unnecessary wear or hazards and are
described below at the frequency with which they should be applied.
Wipe away spilled saline solutions, solvents, acids or
alkaline solutions from outer surfaces immediately to
prevent damage, and wipe with deionized distilled water.
See “Cleaning of the instrument” on page 42.
3
If any surfaces have been contaminated with
biohazardous material, disinfect with a mild sterilizing
solution. See “Instrument care” on page 41.
3
Clean the case of the instrument periodically. See
“Cleaning of the instrument” on page 42.
3
Clean the block wells when necessary. See “Cleaning the
block and wells" on page 43.
3
Clean the heat sink when necessary. See “Cleaning the
heat sink" on page 43.
3
3
Decontaminate the instrument when relocating the
instrument or sending it for service. See
“Decontamination procedure” on page 44.
Service the instrument regularly. See “Cleaning of the
instrument” on page 42 and “Maintaining a system log”
on page 46.
When
sending to
service /
relocating
Yearly
Weekly
Item
When
necessary
Table 5–3. Maintenance checklist
3
3= depending on the laboratory conditions and the use and configuration of the instrument
Instrument care
Thermo Fisher Scientific
For proper instrument care, follow the instructions below.
Thermo Scientific PikoReal Real-Time PCR System User Manual
41
Maintenance
Cleaning of the instrument
●
Always ensure that the electricity supply in the laboratory conforms
to that specified on the type label of the instrument.
●
To guarantee the continuous reliability and accuracy of the
PikoReal Real-Time PCR System, avoid interfering with any of the
optical system components. A misalignment of the light path affects
measurements.
●
Prevent any liquid from entering the instrument.
●
Keep the instrument free of dust and other foreign matter.
●
It is recommended that you clean the case of the instrument
periodically to maintain its good appearance (see “Cleaning of the
instrument” on page 42).
●
In the event of any damage, contact your local Thermo Fisher
Scientific representative for service.
Caution Although the PikoReal Real-Time PCR System is constructed
from high-quality materials, you must immediately wipe away spilled
saline solutions, solvents, acids or alkaline solutions from outer surfaces
to prevent damage and wipe with deionized distilled water. ▲
Caution Keep the instrument block clean to avoid dust and dirt from
entering the measurement chamber. ▲
Cleaning of the
instrument
Clean the instrument regularly as stated below.
1. Turn the power off and unplug the instrument.
2. Wear disposable gloves to protect yourself.
3. Clean the instrument outside and the keypad with a soft cloth
dampened with water.
4. Plastic covers and surfaces can also be cleaned with a mild
laboratory detergent, mild bleach or diluted alcohol solution.
Warning If you have spilt infectious agents on the instrument,
decontaminate the instrument. Refer to “Decontamination procedure”
on page 44. ▲
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Maintenance
Cleaning of the instrument
Caution Painted surfaces can be cleaned with most laboratory
detergents. Dilute the cleaning agent as recommended by the
manufacturer. Do not expose painted surfaces to concentrated acids or
alcohols for prolonged periods of time as damage may occur. ▲
Caution Do not use any solutions containing oxidative chemicals, such
as strong hypochlorites and peroxides or strong bases, on any of the
anodized aluminum surfaces, as this may cause permanent damage to
the finish. ▲
Caution Abrasive cleaning agents are not recommended, because they
are likely to damage the paint finish. ▲
Caution Never use acetone as it will damage the covers. ▲
Caution Do not autoclave any part of this instrument. ▲
Cleaning the block
and wells
To clean the block and wells (Figure 2–2, Figure 2–5 and Figure 2–6),
follow the instructions below.
1. Turn the power off and unplug the instrument.
2. Wear disposable gloves to protect yourself.
3. Clean the block surface at least once a week using a soft cloth or
tissue paper soaked in a mild detergent solution, soap solution or
70% ethanol. Wipe up spills immediately. Do not use
formaldehyde or strong alkalines.
4. Clean the block wells with swabs moistened with water, 95%
ethanol, or a 1:100 dilution of bleach in water.
Warning If you have spilt infectious agents on the block wells,
decontaminate the instrument. Refer to “Decontamination procedure”
on page 44. ▲
Cleaning the heat
sink
Inspect the fins of the heat sink on a regular basis. Dirty heat sinks have
a significantly lower capacity to eject heat.
Clean the fins if they become dirty or covered in dust. Use a cotton
swab, a brush, or compressed air or a vacuum device to remove dust.
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Maintenance
Disposal of materials
Disposal of
materials
Follow laboratory and country-specific procedures for biohazardous or
radioactive waste disposal. Refer to local regulations for the disposal of
infectious material.
Warning The samples can be potentially infectious. Dispose of all used
disposable UTW vessels, strips, and tubes, syringes, disposable tips,
disposable gloves, and so on as biohazardous waste. Be cautious and
always use gloves. ▲
Decontamination
procedure
If you have spilled infectious agents, carry out the decontamination
procedure.
Warning The decontamination procedure should be performed by
authorized and trained personnel in a well-ventilated room wearing
disposable gloves, protective glasses and clothing. ▲
Decontamination should be performed in accordance with normal
laboratory procedures. Any decontamination instructions provided with
the reagents used should be followed.
It is strongly recommended to perform the complete decontamination
procedure before relocating the instrument from one laboratory to
another.
Example of decontaminants:
●
Ethanol 70%
●
Virkon solution 1–3%
●
Glutaraldehyde solution 4%
●
Chloramine T
●
Microcide SQ 1:64
●
Decon 90 min. 4%
To decontaminate the instrument:
1. Wear disposable gloves to protect yourself.
2. Prepare the decontaminant.
3. Switch off the power and disconnect the power supply cable.
4. Empty the drawer.
44 Thermo Scientific PikoReal Real-Time PCR System User Manual
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Maintenance
Packing for service
5. Disinfect the outside of the instrument or to remove stains using a
cloth dampened with 70% ethanol.
6. Place the instrument in a large plastic bag. Ensure that the drawer
door is out.
7. Place a cloth soaked in the prepared decontaminant solution into
the bag. Ensure that the cloth does not come into contact with the
instrument.
8. Close the bag firmly and leave the instrument in the bag for at least
24 hours.
9. Remove the instrument from the bag.
10. After decontamination, clean the instrument using a mild detergent.
11. After performing this decontamination procedure, enclose a signed
and dated Certificate of Decontamination (see Appendix B:
“Certificate of Decontamination”) both inside the transport package
and attached to the outside of the package.
Packing for
service
Thermo Fisher Scientific
To pack the PikoReal Real-Time PCR System for service, follow the
instructions presented below:
●
Inform about the use of hazardous materials.
●
Remove any UTW vessels, tube strips or single tubes before
decontamination. Decontaminate the instrument.
●
Park the instrument. Refer to “Park instrument” on page 38.
●
Use the original packaging for shipping.
●
Pack the instrument according to the packing instructions.
●
Enclose a dated and signed “Certificate of Decontamination” (see
Appendix B) both inside and attached to the outside of the package,
in which you return your instrument (or other items).
●
Enclose the return authorization number (RGA) given by your local
Thermo Fisher Scientific representative.
●
Indicate the fault after you have been in touch with your local
Thermo Fisher Scientific representative or the Thermo Fisher
Scientific technical service department.
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45
Maintenance
Maintaining a system log
Maintaining a
system log
A system log, which includes a short summary of the use, maintenance
procedures, error messages and other information about the use of the
system, is very useful in properly maintaining the system. Refer to
Appendix A: “System Log”. Copy the table as many times as necessary,
but leave the blank original inside the user manual.
Disposal of the
instrument
If the PikoReal Real-Time PCR System is exposed to potentially
infectious chemical samples, toxic or corrosive chemicals or radioactive
chemicals, waste management of the complete instrument must be
carried out to ensure that there is no risk of contamination.
Warning Decontaminate the instrument before disposal. Refer to
“Decontamination procedure” on page 44. ▲
Follow laboratory and country-specific procedures for biohazardous or
radioactive waste disposal.
Dispose of the instrument according to the legislation stipulated by the
local authorities concerning take-back of electronic equipment and
waste. The proposals for the procedures vary by country.
Pollution degree
Method of disposal
2 (see “Safety specifications” on page 48)
Electronic waste
Contaminated waste
(Infectious waste)
Regarding the original packaging and packing materials, use the
recycling operators familiar to you.
For more information, contact your local Thermo Fisher Scientific
representative.
46 Thermo Scientific PikoReal Real-Time PCR System User Manual
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Technical Specifications
Instrument specifications
Chapter 6
Technical Specifications
Instrument
specifications
Thermo Fisher Scientific reserves the right to change any specifications
without prior notice as part of our continuous product development
program (Table 6–4).
Table 6–4. Instrument specifications
Instrument specifications
Dimensions
Size
30 cm (W), 23 cm (D), 31 cm (H) [11.8” (W) x 9.1” (D) x 12.2” (H)]
Weight
9.2 kg [20.3 lbs.] (includes power supply and cord)
Electrical
Power supply
200 W, 24 V, 8.3 A
Line voltage
90–240 VAC
Input current
2.5 A and 4.0 A
Frequency
50–60 Hz
Power failure recovery
Yes (for 24 hours)
Instrument configuration
Block formats
24-well, 96-well (not interchangeable)
Display
Backlit LCD
Communication ports
Ethernet, USB (memory stick only)
Programming
Adjustable ramp rate
Yes
Touchdown
Yes
Typical thermal performance
Max. ramp rate
> 5°C/s heating, > 4.5°C/s cooling
Thermal uniformity
± 0.3°C at 95°C; ± 0.15 at 60°C; ± 0.2 at 72°C
Thermal accuracy
± 0.2°C
Thermal range
4°C and 99.9°C
Typical optical performance
Excitation
5 LEDs
Excitation range
475–640 nm
Detection
CCD
Detection range
520–740 nm
Continued
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Technical Specifications
Safety specifications
Cont.
Instrument specifications
Measuring time
All channels < 13 s
Single channel < 3 s
Filter wavelength range
Channel 1:
Excitation range 475–500 nm
Emission range 520–550 nm
Channel 2:
Excitation range 515–535 nm
Emission range 557–590 nm
Channel 3:
Excitation range 570–590 nm
Emission range 615–650 nm
Channel 4:
Excitation range 600–640 nm
Emission range 666–-740 nm
Channel 5:
Excitation range 475–500 nm
Emission range 520–590 nm
Safety
specifications
In conformity with
the requirements
This section describes the safety specifications for the PikoReal RealTime PCR System.
PikoReal Real-Time PCR System bears the following markings:
Type 5100
CE mark
90–240 Vac, 50/60 Hz
EUT: 24 VDC, 8.33 A
PSU: 90–240 Vac, 50–60 Hz, 2.5 A or 4 A
PikoReal Real-Time PCR System conforms to the following requirements:
2006/95/EC (Low Voltage Directive)
2004/108/EC (Electromagnetic Compatibility Directive, EMC)
2002/96/EC (WEEE Directive)
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Technical Specifications
Safety specifications
The safety specifications are also met under the following environmental
conditions in addition to or in excess of those stated in the operating conditions:
Altitude
Up to 2000 m
Temperature
+5°C to +40°C
Humidity
Maximum relative humidity 80% for temperatures up to
31°C decreasing linearly to 50% relative humidity at 40°C
Mains supply fluctuations
± 10% from nominal
Installation category
(overvoltage category)
II according to IEC 60664-1 (see Note 1)
Pollution degree
2 according to IEC 60664-1 (see Note 2)
Note 1) The installation category (overvoltage category) defines the level
of transient overvoltage which the instrument is designed to withstand
safely. It depends on the nature of the electricity supply and its
overvoltage protection means. For example, in CAT II which is the
category used for instruments in installations supplied from a supply
comparable to public mains, such as hospital and research laboratories
and most industrial laboratories, the expected transient overvoltage is
2500 V for a 230 V supply and 1500 V for a 120 V supply.
2) The pollution degree describes the amount of conductive pollution
present in the operating environment. Pollution degree 2 assumes that
normally only nonconductive pollution, such as dust, occurs with the
exception of occasional conductivity caused by condensation. ▲
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Technical Specifications
Safety specifications
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Troubleshooting Guide
Handling abnormal situations
Chapter 7
Troubleshooting Guide
Note Do not use the instrument if it appears to malfunction. ▲
Handling
abnormal
situations
If there is an abnormal situation during operation, such as fluids spilling
inside the instrument:
1. Switch off the instrument (Figure 2–4 on page 17).
2. Unplug the instrument immediately from the power supply
(Figure 3–10 on page 26).
3. Carry out appropriate corrective measures. However, do not
disassemble the instrument.
4. If the corrective measures taken do not help, contact authorized
technical service or your local Thermo Fisher Scientific
representative.
Power failure
In case of less than a 24-hour power failure, the instrument continues
the interrupted run from the same protocol step it was performing at
the time of the interruption.
If the protocol was in the melting curve phase (see the PikoReal Software
User Manual), the instrument continues the run from the start of the
melting curve protocol.
The instrument gives a warning code dependent of the length of the
power failure (W1 or E1, see Table 7–5).
Warning and
error codes
Warnings inform about possible unwanted events occurring during the
protocol execution (Table 7–5). The protocol is not stopped, but the
event might have some effect on the results.
Errors inform about a failure occurring during the protocol execution
(Table 7–6). All errors, except E1, cause the protocol to stop and all
heating units to be turned off.
Thermo Fisher Scientific
Thermo Scientific PikoReal Real-Time PCR System User Manual
51
Troubleshooting Guide
Warning and error codes
When an error is detected, the current operation is aborted. After an
error, it is best to restart from the beginning after the problem has been
fixed. When an error occurs, the error code and the suggested actions
are presented. You can restore the data of the last run in event of
technical problems.
Press [C] to clear the warning or error message.
Table 7–5. Warnings
Code
Symbolic name
Description
Action text
W1
POWER FAILURE
This warning is set if the power was
lost during protocol execution for less
than 24 h and the protocol was
recovered.
CHECK LOGFILES
W8
BLOCK TEMP HIGH
This warning is set if the block
temperature is over 112°C.
W9
LID TEMP HIGH
This warning is set if the lid
temperature is over 115°C.
W10
SINK TEMP HIGH
This warning is set if the sink
temperature is over 75°C.
W27
PROTOCOL STOPPED
This warning is set if the protocol is
stopped before the end of the protocol.
W28
CAMERA FAILURE
This warning is set if the first camera
plate read command fails (only on RT).
W29
PROTOCOL PAUSED
This warning is set if the protocol is
paused during the protocol run.
CHECK LOGFILES
W30
CYCLES ADDED
This warning is set if extra cycles are
added to a protocol loop.
CHECK LOGFILES
W31
STEP SKIPPED
This warning is set if the protocol step
is skipped (NEXT).
CHECK LOGFILES
W32
LID TEMP OVERRIDDEN
This warning is set if the lid
temperature is overridden from the
service menu.
CHECK LOGFILES
CHECK THAT THE
HEATSINK IS CLEAN
Table 7–6. Errors
Code
Symbolic name
Description
Action text
E1
POWER FAILURE
This error is set if the power was lost
during protocol execution for over 24 h.
CHECK LOGFILES
E3
BLOCK TEMP.
This error is set if the block
temperature is over 115°C.
CONTACT SUPPORT
E4
LID TEMP.
This error is set if the lid temperature
is over 125°C.
CONTACT SUPPORT
E5
SINK TEMP.
This error is set if the sink temperature
is over 85°C.
ENSURE THAT THE
AIRFLOW TO THE FAN
IS NOT OBSTRUCTED.
IF PROBLEM PERSIST
CONTACT SUPPORT
Continued
52 Thermo Scientific PikoReal Real-Time PCR System User Manual
Thermo Fisher Scientific
Troubleshooting Guide
Warning and error codes
Cont.
Code
Symbolic name
Description
Action text
E11
CALIB. TABLE
This error is set if any calibration
values or the calibration derived from
them is corrupt.
REBOOT
INSTRUMENT. IF
PROBLEM PERSISTS
CONTACT SUPPORT
E12
SINK TEMP. LOW
This error is set if the sink temperature
is below -20°C.
WAIT FOR THE
INSTRUMENT TO
REACH OPERATING
TEMP. IF PROBLEM
PERSISTS CONTACT
SUPPORT
E13
BLOCK TEMP. LOW
This error is set if the block
temperature is below -20°C.
WAIT FOR THE
INSTRUMENT TO
REACH OPERATING
TEMP. IF PROBLEM
PERSISTS CONTACT
SUPPORT
E14
LID TEMP. LOW
This error is set if the lid temperature
is below -20°C.
WAIT FOR THE
INSTRUMENT TO
REACH OPERATING
TEMP. IF PROBLEM
PERSISTS CONTACT
SUPPORT
E15
BLOCK TEMP. SHIFT
This error is set if the block
temperature changes more than 10°C
during a 500 ms interval.
REBOOT
INSTRUMENT. IF
PROBLEM PERSISTS
CONTACT SUPPORT
E17
LID TEMP. SHIFT
This error is set if the lid temperature
changes more than 20°C during a
500 ms interval.
REBOOT
INSTRUMENT. IF
PROBLEM PERSISTS
CONTACT SUPPORT
E18
LID POS. FAILURE
This error is set if the lid encounters
pressure too early. In case of Piko 24,
this makes it incompatible with
doomed caps.
RETRY. IF PROBLEM
PERSISTS SEE
MANUAL
E21
BLOCK TEMP. CHANGE
This error is set if the block is heated
with more than a 6 A current for 5 sec
and the change in the block
temperature is not bigger than 1°C.
CONTACT SUPPORT
E22
LOG FILE
The log files have been corrupted and
the log file memory has been cleared.
REBOOT
INSTRUMENT. IF
PROBLEM PERSISTS
CONTACT SUPPORT
E31
CAMERA FAILURE
This error is set if the camera plate
read command fails twice.
REBOOT
INSTRUMENT. IF
PROBLEM PERSISTS
CONTACT SUPPORT
E32
RT-UNIT FAILURE
This error is set if the connection to the
RT unit fails.
REBOOT
INSTRUMENT. IF
PROBLEM PERSISTS
CONTACT SUPPORT
Continued
Thermo Fisher Scientific
Thermo Scientific PikoReal Real-Time PCR System User Manual
53
Troubleshooting Guide
Warning and error codes
Cont.
Code
Symbolic name
Description
Action text
E33
SELF TEST FAILURE
This error is set if the RT self test fails.
The self test is run at every boot and
protocol start and it checks that the
camera is found, the filter is initialized
and the plate is configured (for a final
tested instrument).
REBOOT
INSTRUMENT. IF
PROBLEM PERSISTS
CONTACT SUPPORT
E35
LID TEMP. CHANGE
This error is set if the lid is active, the
temperature difference to the target is
more than 20°C and the response to a
10 seconds heating is less than 2°C.
CONTACT SUPPORT
E36
STRAIN GAUGE FAILURE
This error is set if the lid pressure
sensor is broken.
CONTACT SUPPORT
E37
LID OPTO FAILURE
This error is set if the lid position
sensor is unable to detect the home
position.
RETRY. IF PROBLEM
PERSISTS SEE
MANUAL
E40
DRAWER FAILURE
This error is set if the drawer is
opened, but the drawer opto state is
not changed in 100 ms.
CONTACT SUPPORT
54 Thermo Scientific PikoReal Real-Time PCR System User Manual
Thermo Fisher Scientific
Ordering Information
PikoReal
Chapter 8
Ordering Information
Contact your local Thermo Fisher Scientific representative for ordering
and service information (Table 8–7 through Table 8–11).
PikoReal
List of spare
parts and
accessories
Thermo Fisher Scientific
Table 8–7. Instrument catalog number
Code
Item
TCR0024
PikoReal Real-Time PCR System, 24-well
TCR0096
PikoReal Real-Time PCR System, 96-well
Table 8–8. Codes for spare parts and accessories
Code
Item
Qty
N11471
PikoReal Real-Time PCR System User Manual
1
N12076
PikoReal Software User Manual
1
N12119
PikoReal Software for PikoReal Real-Time PCR System CD
1
EA12101M-240-VAN
CMP power supply EA12101M-240-VAN
1
5180220
Power cable (UK)
1
1210550
Power cable (Euro)
1
1210520
Power cable (USA/Japan)
1
N08421
Power cable (China)
1
N12094
Ethernet cable
1
TCR0096DEMO
PikoReal 96 demo kit (1 plate)
1
TCR0024DEMO
PikoReal 24 demo kit (1 plate)
1
TCR0096VAL
PikoReal 96 validation plate
1
TCR0024VAL
PikoReal 24 validation plate
1
Thermo Scientific PikoReal Real-Time PCR System User Manual
55
Ordering Information
List of consumables
List of
consumables
Table 8–9. Codes for consumables
Code
Item
Quantity
SPL0240
Thermo Scientific Piko PCR Plate, 24-well, clear
200 plates
SPL0241
Thermo Scientific Piko PCR Plate, 24-well, white
200 plates
SPL0960
Thermo Scientific Piko PCR Plate, 96-well, clear
200 plates
SPL0961
Thermo Scientific Piko PCR Plate, 96-well, white
200 plates
SFR0241
Thermo Scientific Piko 24-well PCR Plate Frame, white
50 frames
SFR0961
Thermo Scientific Piko 96-well PCR Plate Frame, white
50 frames
TCS1080
Thermo Scientific flat 8-cap strips for Piko PCR Plates,
optically clear
120 strips
AB-0812
Clear Seal Diamond Heat Sealing Film
100 seals
TUC0010
Thermo Scientific Piko UTW individual tubes, clear
tube/clear cap
960 tubes
TUC0011
Thermo Scientific Piko UTW individual tubes, white
tube/clear cap
960 tubes
TUC0080*
Thermo Scientific Piko UTW 8-tube strips w/attached
caps, clear
250 strips
TUC0081*
Thermo Scientific Piko UTW 8-tube strips w/attached
caps, white
250 strips
ASF0020
Thermo Scientific sealing film, Piko PCR Plate size
400 seals
AB-0771
Low Profile Thermo Strip, clear
0.2 ml low profile thin walled 8 tube strips
120 strips
AB-0771/w
Low Profile Thermo Strip, white
0.2 ml low profile thin walled 8 tube strips
120 strips
AB-0866
Ultra Clear Cap Strips
120 strips
* Not available in US.
56 Thermo Scientific PikoReal Real-Time PCR System User Manual
Thermo Fisher Scientific
Ordering Information
List of reagents
List of reagents
Table 8–10 shows examples of reagents available for the PikoReal. For
more information and other available products, refer to
http://www.thermoscientific.com/pikoreal.
Table 8–10. List of reagents
SYBR® Green chemistry
K0251/ K0252
Maxima™ SYBR Green qPCR Master Mix (2X), ROX Solution provided
F-415S/L
DyNAmo™ Flash SYBR Green qPCR Kit
F-416S/L
DyNAmo™ ColorFlash SYBR Green qPCR Kit
Probe based chemistry
K0261/K0262
Maxima™ Probe qPCR Master Mix (2x), ROX Solution provided
F-455S/L
DyNAmo™ Flash Probe qPCR Kit
F-456S/L
DyNAmo™ ColorFlash Probe qPCR Kit
SNP Genotyping (based on hydrolysis probes)
F-480S/L/XL
DyNAmo™ SNP Genotyping Master Mix
®
Solaris qPCR Gene Expression Assays (human and mouse)
AX-XXXXXX-XX-0X00*
*Catalog number is product-specific.
Please refer to
www.thermoscientific.com/solaris to
select the appropriate assay
Each pre-designed Solaris® qPCR Assay consists of a
single probe (200 nM) and primer pair (800 nM each)
specific to your target gene. Each assay is designed to
a consensus sequence that covers all known splice
variants of the target gene. The sequence information
is provided.
Solaris® qPCR Gene Expression Master Mix
Plus ROX vial / including ROX / including Low ROX
AB-4350 / AB-4351 / AB-4352
Solaris® RNA Spike Control Kit
K-002200-C1-XXXX
Package size (XXXX):
100/200/400/1000 x 25µL rxns
cDNA Synthesis Kits for qPCR
K1641/ K1642
Maxima™ First Strand cDNA Synthesis Kit for RT-qPCR
AB-1453/A, AB-1453/B
Verso™ cDNA Synthesis Kit (with Solaris® qPCR Master Mixes)
List of heat
sealers
Thermo Fisher Scientific
Table 8–11. List of heat sealers
Code
Item
AB-0384/110
Thermo Scientific ALPS 25 Manual Heat Sealer, 110 V
AB-0384/220
Thermo Scientific ALPS 25 Manual Heat Sealer, 220 V
AB-1443
Thermo Scientific ALPS 50 V Microplate Heat Sealer
Thermo Scientific PikoReal Real-Time PCR System User Manual
57
Ordering Information
List of heat sealers
58 Thermo Scientific PikoReal Real-Time PCR System User Manual
Thermo Fisher Scientific
System Log
Appendix A
System Log
Instrument name/number:
User
Date
Comments
PHOTOCOPIABLE
Thermo Fisher Scientific
Thermo Scientific PikoReal Real-Time PCR System User Manual 59
System Log
60
Thermo Scientific PikoReal Real-Time PCR System User Manual
Thermo Fisher Scientific
Certificate of Decontamination
Appendix B
Certificate of Decontamination
Name:
Address:
Tel./Fax:
Instrument:
Serial no.:
A) I confirm that the returned items have not been contaminated by body fluids, toxic,
carcinogenic or radioactive materials or any other hazardous materials.
B) I confirm that the returned items have been decontaminated and can be handled without
exposing the personnel to health hazards.
Materials used in the unit: Chemicals +
Biological •
Radioactive *)
Specific information about contaminants:
Decontamination procedure1:
Date and place:
Signature:
Name (block capitals):
*) The signature of a Radiation Safety Officer is also required when the unit has been used with
radioactive materials.
This unit is certified by the undersigned to be free of radioactive contamination.
Date and place:
Signature:
Name (block capitals):
PHOTOCOPIABLE
1
Thermo Fisher Scientific
Please include decontaminating solution used.
Thermo Scientific PikoReal Real-Time PCR System User Manual 61
Certificate of Decontamination
62
Thermo Scientific PikoReal Real-Time PCR System User Manual
Thermo Fisher Scientific
Glossary
CCD Charge-coupled device.
intercalator In biochemistry, intercalator is a type
of molecule that binds to DNA and inserts itself
into the DNA structure.
LED Light-emitting diode.
MAC Short for Media Access Control address, a
hardware address that uniquely identifies each
node of a network.
PCR Polymerase chain reaction.
qPCR Quantitative real-time PCR.
RFU Relative fluorescence unit.
UI User interface.
UTW Ultra-thin wall.
Thermo Fisher Scientific
Thermo Scientific PikoReal Real-Time PCR System User Manual 63
Glossary:
64 Thermo Scientific PikoReal Real-Time PCR System User Manual
Thermo Fisher Scientific
Index
A
E
accessories, 23, 24, 55, 58
accuracy, 2, 42, 47
air, 16, 24, 25, 43
exhaust, 16
flow, 25
intake, 16
allelic discrimination, 11
application, 2, 5, 11, 12, 13, 14, 18
automatic lid, 12
emission, 48
environmental requirements, 24, 25
error, 17, 27, 29, 46, 51, 52, 53, 54
codes, 27, 51, 52
messages, 27, 46, 52
Ethernet connection, 12, 24, 26, 30, 55
external power supply, 2, 12, 17, 18, 24, 25, 26, 44, 47, 51,
55
F
B
block, 5, 11, 12, 16, 18, 19, 20, 32, 36, 41, 42, 43, 47, 52,
53, 61
format, 11, 12, 18, 47
wells, 41, 43
C
calibration, 21, 53
CCD camera, 12
clock, 34, 35, 37
communication ports, 47
consumables, 3, 13, 18, 20, 32, 56
cycle, 33
D
data port, 26
decontamination, 41, 42, 43, 44, 45, 46, 61
Certificate of, 45, 61
procedure, 41, 42, 43, 44, 45, 46, 61
diagnostics, 2, 34, 35, 36, 37
display, 29, 33, 34, 35, 36, 37, 47
disposal of instrument, 46
disposal of materials, 44
DNA analysis, 13
drawer, 16, 36, 44, 45, 54
dye calibration, 21
file, 33, 53
fluorescence, 13, 63
intensity, 13
fluorogenic, 13
folder, 30, 33
USB, 30
G
glossary, 63
H
heat, 11, 12, 18, 19, 21, 25, 41, 43, 56, 57
sink, 11, 12, 25, 41, 43
heated lid, 5, 12
heating, 29, 47, 51, 54
how to clean the block wells, 41, 43
how to clean the heat sink, 41
how to decontaminate the instrument, 41, 42, 43, 44, 45, 46,
61
how to ensure startup, 29
how to install the PikoReal, 23, 25, 27, 49
how to maintain the instrument, 41
how to pack for service, 23, 45
how to switch on, 29
how to unpack, 23
I
installation, 3, 23, 25, 27, 29, 49
Thermo Fisher Scientific
Thermo Scientific PikoReal Real-Time PCR System User Manual
65
category, 49
intended use, 3, 13
intercalator, 63
power supply, 2, 12, 17, 18, 24, 25, 26, 44, 47, 51, 55
protocol, 27, 29, 30, 31, 32, 33, 34, 38, 40, 51, 52, 54
Q
L
LED, 12, 17, 29, 32, 63
function, 17, 29
lid, 5, 12, 29, 32, 34, 35, 36, 52, 53, 54
logfiles, 34, 38, 52, 53
qPCR, 11, 13, 20, 57, 63
reagents, 13, 20
technology, 13
quantification, 11, 13
R
M
maintenance, 18, 23, 25, 41, 42, 46
checklist, 41
measurement, 42
chamber, 42
melt curve, 11
melting, 11, 51
multiplexing, 12, 13
N
network, 26, 34, 37, 63
O
operational, 27
check, 27
optical, 12, 19, 42, 47
filter, 12
performance, 47
system, 12, 42
ordering information, 55
P
packing, 23, 24, 45, 46
instructions, 24, 45
list, 23
materials, 46
park instrument, 34, 38, 45
PCR, 3, 4, 5, 11, 12, 13, 16, 17, 18, 19, 20, 21, 23, 24, 25,
26, 27, 29, 31, 40, 42, 45, 46, 48, 55, 56, 63
cycle, 13
method, 13
product, 13
PIKO, 35
plate, 11, 18, 19, 31, 32, 52, 53, 54, 55, 56
drawer, 16, 36, 44, 45, 54
66 Thermo Scientific PikoReal Real-Time PCR System User Manual
ramp rate, 47
reagents, 11, 12, 21
RFU, 63
RT, 35, 52, 53, 54, 57
running a protocol, 26, 30
S
S/N, 35
sealer, 21, 57
sensors, 35, 36
service, 5, 23, 27, 34, 36, 38, 41, 42, 45, 51, 52, 55
shutdown, 40
sink, 11, 25, 36, 41, 43, 52, 53
spare parts, 3, 55
specifications, 3, 46, 47, 48, 49
safety, 46, 48, 49
startup, 29
Status screen, 29, 32, 33, 34
step, 33, 34, 51, 52
switching on, 29
symbols, 4
system info, 34, 35
system log, 41, 46
T
Tools menu, 34, 35, 36, 37, 38, 39
transport, 23, 24, 39, 45
lock, 39
package, 24, 45
troubleshooting, 3, 17, 29, 36, 51
tubes, 12, 19, 31, 32, 40, 44, 45, 56
type, 4, 26, 27, 31, 35, 42, 48, 63
type label, 4, 27, 42
Thermo Fisher Scientific
U
W
uniformity, 11, 47
USB, 12, 26, 29, 30, 31, 34, 39, 40, 47
user interface, 17, 29, 63
UTW, 2, 11, 13, 19, 20, 32, 40, 44, 45, 56, 63
warning codes, 26, 51, 52
warranty, 5, 23
wavelength, 48
range, 48
Thermo Fisher Scientific
Thermo Scientific PikoReal Real-Time PCR System User Manual
67
Notes
68 Thermo Scientific PikoReal Real-Time PCR System User Manual
Thermo Fisher Scientific
Plesmanlaan 1d
Thermo Fisher Scientific
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Ratastie 2, P.O. Box 100
The Netherlands
T. +31 (0)71 568 10 00
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T. Belgium: 0800 71640
Finland knowledge F. +31 (0)71 568 10 10
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N11471
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