Thermal cycler / Thermocycler 0612E+D Creating the future together. Short Manual peqSTAR Thermocylcer CONTENTS 1 Safety instructions 2 License Notice 3 System Overview 4 Installation 4.1 Content of delivery 4.2 Installing the thermal cycler 4.3 Operation of the thermal cycler 5 Cycler Operation 5.1 Start screen = 'Run' 5.2 Running stored programs ('Run'): 5.3 Incubation mode ('Run') 5.4 Create or edit PCR programs ('Programs') 5.5 Further dialogues: GLPs, Diagnostics, System 5.6 Further questions…? 6 Technical Service and Ordering Information 7 Technical data 8 Maintenance and repair of the Thermal cycler 8.1 Cleaning, maintenance and repair of the thermal cycler 8.2 Temperature check-up 8.3 Calibration 9 Warranty 10 Disposal Instruction 2 2 3 3 3 3 3 4 4 4 5 5 8 9 10 11 12 12 12 12 13 13 A more detailed instruction manual in English or German is available for download on our website www.peqlab.com. PEQLAB_v0612D 1 Short Manual peqSTAR Thermocylcer 1 SAFETY INSTRUCTIONS Before the first use of the thermal cycler please read this instruction manual entirely. Special notice should be taken of the following Caution of dangerous voltage Please ensure the voltage indicated on the device exactly matches your local electrical supply Caution of dangerous explosive material Explosive or reactive material mustn’t be heated in the thermal cycler. Caution - liquids Ensure that no liquids can enter the device. Caution - hot surfaces The thermal plate and the heated lid quickly reach temperatures above 50 °C. There is danger of burning! Keep the lid closed until the temperatures reaches 30 °C or less. Only use materials (tubes, tube caps, plates and sealing films) which are heat resistant to 120 °C Caution – environment The ventilation of the device must not be covered. 2 LICENSE NOTICE This instrument is licensed for research and development and for uses other than human in vitro diagnostics under one or more of the following patens of Applera Corporation: U.S. Patent Nos. 5,656,493, 5,038,852, 5,333,675, 5,475,610 (claims 1-159 and 164-166), 6,703,236 (claims 7-10) and 7,238,517 or corresponding claims in their non-U.S. counterparts. No right is conveyed expressly, by implication or by estoppels under any other patent of Applera, including but not limited to U.S. Patent No. 6,814,934 and its non-U.S. counterparts, which describe and claim thermal cyclers capable or real-time detection. PEQLAB_v0612D 2 Short Manual peqSTAR Thermocycler 3 The peqSTAR thermal cycler is the ideal PCR device for all molecular biological and biochemical laboratories working in the field of basic research or routine diagnostic. Six 250 °C HTR (High Temperature Range) Peltier elements allow quick changes in temperature up to 5 °C/sec (max). The individual monitoring and controlling of each of the six Peltier elements equipped with 'Long Life Technology' ensure the tightest correlation of target and actual temperature across the entire thermal plate leading to highest reproducibility of the results. The peqSTAR thermal cycler is equipped with a touch-sensitive, graphic colored TFT display of the newest generation, allowing operation of the device by simply touching symbols and graphic elements on the screen. This leads to easy and intuitive handling and programming of the device. Additionally a mouse can be connected via USB. Further-more an Ethernet port is available to connect the device with the Microsoft Windows® network. It is therefore possible to use an external server for archiving programs and GLP reports which will be available globally in the network (e.g. for other thermal cyclers). Transfer of data can be done by using commercially available USB memory sticks enabling a nearly unlimited saving space for programs and GLP reports, which can be printed using a printer connected to the thermal cycler via the network. Notice: 4 4.1 The USB ports support only standard cable mice and standard USB sticks. The maximum acceptable length of the connecting cable of the port is 3 m! INSTALLATION Content of delivery 1 Thermal cycler 1 Power cord 1 Short manual (A detailed instruction manual in English or German is available for download on our website www.peqlab.com.) 1 CD including PC-software 'peqSTAR Protocol Editor' Please check delivery for completeness and transport damage upon arrival. If any transport damage is noticed, contact your distributor or manufacturer immediately. NOTICE: IF ANY DAMAGE OF THE DEVICE SHOULD BE FOUND DO NOT USE THE DEVICE! 4.2 Installing the thermal cycler Check the packaging for any transport damage. Remove the packaging and place the thermal cycler onto a solid surface. The device should not be exposed to direct sunlight. There should be enough space to make sure that the ventilator grill on the front and back are not covered and sufficient ventilation is provided. There should be a distance of at least 25 cm to the wall or the next device. Two thermal cyclers should not be placed back-to-back. Room temperature between 4 °C und 25 °C is advised. All original packaging should be stored in case a need arises to return the instrument to the supplier. 4.3 Operation of the thermal cycler Before starting up the thermal cycler for the first time make sure that the voltage range of the device is suitable to the electricity supply. Plug in the mains plug of the thermal cycler to the electricity supply. The outlet and the power switch are located at the back of the device. The serial number is written onto a typed label at the bottom of the device. The device should only be used by authorized staff. To ensure this, a user administration with appointing rights for single users is available in the device. PEQLAB_v0612E 3 Short Manual peqSTAR Thermocycler 5 CYCLER OPERATION The operation of the thermal cycler can be run by the internal touch screen, either using the touch sensitive surface of the display or by using a mouse connected to a free USB port. If the user has to enter numbers or text the corresponding field has to be touched on the touch screen or clicked with the mouse. An appropriate keyboard will appear on the screen according to the expected entry. 5.1 Start screen = 'Run' The last ran PCR program is displayed in the 'Run' dialogue and can directly be restarted with the button 'Start PCR'. 5.2 Running stored programs ('Run'): 1. Open the selection dialogue in the 'Run' dialogue with the button 'Select Program'. 2. Select the desired program in the selection dialogue and load to the 'Run' dialogue with the button 'Open'. 3. The loaded program is started by using the 'Start' button. PEQLAB_v0612E 4 Short Manual peqSTAR Thermocycler 5.3 Incubation mode ('Run') Use the button 'Run Incubation' in the 'Run' dialogue to set the device into the incubation mode. The temperature for the thermal plate and lid can be set manually. The clock shows the time passed by in the incubation mode. 5.4 Create or edit PCR programs ('Programs') 1. To create or edit PCR programs go to the 'Programs' dialogue. 2. By touching the 'Programs' dialogue the first time during the session the selection dialogue shows up where a new program can be set over 'New' or an existing program can be opened over 'Open' to edit. 3. On the left side of the program editor all available commands are displayed, on the right side of the editor the list of commands (Steps) the program is composed of is shown. After selecting a program command out of the list in the left window it can be inserted into the desired place via the '+' arrow. 4. Via double click or the button 'Edit' parameters of single steps could be changed later. 5. Use the 'Save' button to save the program. 6. For even faster programming the peqSTAR thermocycler comes with a selection of template programs, which could easily be edited and saved under a new program name. PEQLAB_v0612E 5 List of available commands Program name Adding and removing program steps Program window Short Manual peqSTAR Thermocycler Available commands: Close Lid Loop Locks the lid avoid accidental opening. Combines Open Loop, Close Loop, Temperature, Ramp and Gradient. Open Lid Pause Unlocks the lid in case that the lid has been locked in a previous program step. Interrupts the program temporarily at the definable temperature. Heat Lid Store Turns the lid heating on/off. Holds the definable temperature, turns off the heated lid automatically. Temperature Open Loop Temperature step for definable time, Temperature and Time Incr./Decr. function. Manual loop beginning. Ramp Close Loop Target temperature is reached with definable ramp rate. Manual loop ending. Gradient Realizes different temperatures at the same time for protocol optimization. Sample program Create a new program with 'New'. Enter a program name and confirm with OK to access the programming functions. 01 Heated Lid on, temperature 110 °C Heat Lid A 'Heat Lid' step is inserted in each program by default as first step. This step can be edited or deleted later. The standard settings could be changed under 'System' => 'Firmware' => 'Settings'. To enter another program step select the desired command out of the list of the available command and add it to the program window with the '+' arrow. Thereupon the corresponding parameters will be asked. 02 Denaturation at 95 °C for 2 minutes Temp 03 07 Loop 04 Open the 'Loop' function → Enter number of cycles – 30 cycles → 3 x Continue with 'Temp' → Terminate 'Loop' function with 'Exit Loop' Denaturation at 95 °C for 30 seconds Temp 05 Annealing at 65°C in the 1st cycle for 30 seconds 'Touchdown'-PCR via Temperature-'Decrement'. The annealing temperature of 65 °C in the 1st cycle is reduced by 0.4 °C each cycle. This leads to high specifity in the first cycles and PEQLAB_v0612E 6 Short Manual peqSTAR Thermocycler Temp high amplification efficiency in the later cycles. Elongation at 72 °C for 1 minute in the 1st cycle 06 Temp 07 Via Time -'Increment' the elongation time is prolonged by 5 s each cycle. The incremental prolongation of the elongation time is useful with high numbers of cycles (>30), when the Polymerase activity declines. Terminate 'Loop' function with 'Exit Loop'. Alternative programming of steps 03 – 07: 03 Open program loop – 30 cycles. Loop Open 04 Denaturation at 95 °C for 30 seconds Temp Annealing at 65°C in the 1st cycle for 30 seconds 05 Temp 'Touchdown'-PCR via Temperature-'Decrement'. The annealing temperature of 65 °C in the 1st cycle is reduced by 0.4 °C each cycle. This leads to high specifity in the first cycles and high amplification efficiency in the later cycles. Elongation at 72 °C for 1 minute in the 1st cycle 06 Temp 07 Via Time -'Increment' the elongation time is prolonged by 5 s each cycle. The incremental prolongation of the elongation time is useful with high numbers of cycles (>30), when the Polymerase activity declines. Terminate program loop with 'Close Loop'. Close Loop 08 Final elongation at 72 °C for 5 minutes The final elongation assures the complete synthesis of all amplificats. Temp The ramp starts from the last programmed temperature (here 72 °C) and decreases temperature to the set temperature (here 8°C) at a constant rate. 09 Cools down to 8 °C with a ramp rate of 1.0 °C/s Ramp 10 The reduction of the cooling rate at this step of the program is not necessary, but should only be an example. It might make sense e.g. for quantitative annealing of complementary oligo nucleotide or at annealing steps in the first cycles, when primer annealing should be very efficient because of low template amount (e.g. after cDNA synthesis). 8 °C forever, lid heat is turned off Store PEQLAB_v0612E In the 'Store' step the lid heat is turned off automatically. This avoids that lid heat and Peltier elements of the block work against each other. Especially at longer incubation times at low temperature this increases the life time of the Peltier elements and reduces the power consumption. 7 Short Manual peqSTAR Thermocycler 5.5 Further dialogues: GLPs, Diagnostics, System GLPs: Displaying and organizing GLP (Good Laboratory Practice) reports created by the thermal cycler. This option is mainly for labs who need to save GLP reports for documentation purpose. Diagnostics: Monitoring the actual temperatures of each of the six Peltier elements during a run. In suspicion case of an instrument malfunction have a look at the Peltiers’ temperature profile. System: Configuration of the system and changing of the general settings of the device. PEQLAB_v0612E 8 Short Manual peqSTAR Thermocycler Elements of the system dialogue: Change User Clock Change from one user to another. Setting up date and time. User Accounts Hide/Show Curser Define user and user groups (and their access rights). Hide or show the cursor on the screen. Sounds Set up a printer path for an existing network printer. Setting of the user call sounds (Pause/Store, Program End, Open Lid Call) Protocol (OFF/ON) Network Activate/deactivate the protocol function (Internal communication for failure diagnosis) Network settings for network integration of the peqSTAR. Firmware Display Settings Display, administrate and update software (Control/Power Unit, Thermo block) Change settings of the screen display. Hardware Display hardware versions of Control Unit, Power Unit and Thermo block 5.6 Further questions…? Direct help : Use this button to activate (deactivate) the direct help mode. If the mouse pointer is visible a question mark appears alongside. By touching a function button or other dialogue element the corresponding help window will appear. A more detailed instruction manual in English or German is available for download on our website www.peqlab.com. PEQLAB_v0612E 9 Short Manual peqSTAR Thermocycler 6 TECHNICAL SERVICE AND ORDERING INFORMATION Should you have any questions about the peqSTAR thermocycler, please visit www.peqlab.com to find the respective contact person. This short manual is delivered with the following products: Product peqSTAR 96 Universal peqSTAR 96 Universal Gradient peqSTAR 96 HPL peqSTAR 96 HPL Gradient peqSTAR 384 HPL peqSTAR in situ PEQLAB_v0612E Cat. No. with 96 well Universal block and standard lid for 96 x 0.2 ml tubes, 96 well PCR plates or 48 x 0.5 ml tubes with flat caps with 96 well Universal Gradient block and standard lid for 96 x 0.2 ml tubes, 96 well PCR plates or 48 x 0.5 ml tubes with flat caps with 96 well block and High Pressure Lid for 96 x 0.2 ml tubes or 96 well PCR plates with 96 well Gradient block and High Pressure Lid for 96 x 0.2 ml tubes or 96 well PCR plates with 384 well block and High Pressure Lid for 384 well PCR plates with in situ block for up to four microscope slides and integrated buffer reservoir 10 95-95002 95-96002 95-95002-HPL 95-96002-HPL 95-950384-HPL 95-950FLAT Short Manual peqSTAR Thermocycler 7 TECHNICAL DATA General characteristics 6 Peltier elements with Long-Life-technology and 6 control circuits with Pt 1000 temperature sensors Maximal heating and cooling rate: 5 °C/s Block uniformity (at 72 °C): ± 0.25 °C Temperature range thermoblock: 4 to 105 °C Control accuracy thermoblock: ± 0.1 °C Variable ramping: 0.1 to 2.0 °C/s Increment/Decrement time: 0:01 to 9:59 minutes Increment/Decrement temperature: 0.1 to 9.9 °C Lid heating temperature range: 70 to 120 °C Programmable Lid locking mechanism 4 x USB, 1 x Ethernet interfaces (MS Windows®), maximal length of power cord 3 m! Note: USB interfaces only support standard USB sticks, cord mouse and keyboards! Dimensions (B x H x D): 29 x 31 x 36 cm Weight incl. block: 13 kg Electrical power supply: 90 – 264 V AC, 50 – 60 Hz, 850 VA Fuse: 10 A time lag Gradient feature (optional) Maximal gradient range over 12 columns: 39.8 °C (± 19.9 °C) Temperature range Gradient: 35 to 105 °C Gradient accuracy: ± 0.1 °C User interface/functions Touch sensitive TFT-Display (1/4 VGA, Graphic, 16 colours), operation via USB mouse possible Direct help function, Tm- and elongation time calculator Internal flash memory for 1000 typical PCR programs in free configurable folders/subfolders Unlimited number of programs via network PC or USB memory stick Free computer Software for the generation of PCR programs on the computer Up to 99 steps per program Password protected user accounts with variable access rights GLP reports for complete recording of all runs Quickstart function of the last used program Automatic restart after power failure, Power-Fail-Denaturation, Instant Incubation Interchangeable block options 96 Well universal block and lid heating with automatic height adaptation for 96 x 0.2 ml tubes, 96 Well PCR plates or 48 x 0.5 ml tubes with flat cap 96 Well block and high pressure lid (HPL, 100 – 250 N) for the leak-proof closure of 96 Well plates 384 Well block and high pressure lid (HPL, 100 – 250 N) for the leak-proof closure of 384 Well plates In situ block with integrated buffer reservoir for up to 4 microscope slides PEQLAB_v0612E 11 Short Manual peqSTAR Thermocycler 8 8.1 MAINTENANCE AND REPAIR OF THE THERMAL CYCLER Cleaning, maintenance and repair of the thermal cycler For safety reasons the device must be switched off and the mains plug pulled before cleaning is carried out! The surface of the thermal cycler can be wiped off with a damp cloth or ethanol. Do not use aggressive or scouring cleaners or organic solvents for cleaning. The device should be protected from aggressive chemicals. Make sure that no liquid penetrates into the interior of the device. If sample liquids penetrates into the drill holes of the thermal cycler clean them immediately with mild soap followed by distilled water. You can use mild disinfectants for decontamination. Maintenance (temperature check-up see 13.2) should be carried out regularly to detect possible technical failures. The electric fuses are placed at the back of the device between the power switch and the mains plug. After pulling off the main plugs and pushing upwards a safety catch the fuses can be changed. Use only fuses with correct values (indications about the fuse type are placed at the back of the device). The device may only be opened by a qualified specialist. Unauthorized work on the device voids the warranty. All kinds of repairs may only be carried out by authorized persons using original replacement parts. The replacement of single components of the device should not be done by the user but exclusively by authorized specialists. Therefore the device must be sent to the manufacturer. The surface of the device should be decontaminated with a lint free cloth soaked with 70 % ethanol before sending. The drill holes of the thermoblock shall be cleaned with a 1.5 % hypochlorite solution with the help of a cotton bud. Please fill in and sign the decontamination certificate and send it to the manufacturer together with the device. 8.2 Temperature check-up Maintenance of the temperature at regular intervals (e.g. annually) is recommended. The thermometer for use should be equipped with wired sensing devices (e.g. Pt1000) and should have an accuracy of measurements of 1/10 class B (∆T = ±0.03 °C) in the range of 0 to 100 °C. While processing the temperature control the room temperature should be 25 °C at maximum. 8.3 Calibration If there is a reasonable suspicion that the device has deviated from the calibration range, because the variations of the temperature check-up are considerably out of the acceptable tolerance, a new calibration must be carried out by the manufacturer. Please contact your local distributor. PEQLAB_v0612E 12 Short Manual peqSTAR Thermocycler 9 WARRANTY The period of warranty for the product at hand is 24 month from date of purchase. The warranty does not apply for defects caused by incorrect, improper or inappropriate use. The customer has to check up on obvious defects promptly upon receipt of the device and send a written report. Otherwise the claim for warranty expires. The manufacturer reserves the right to carry out changes to the thermal cycler or to the technical documentation of the thermal cycler on the basis of product development or improvement without prior notice. 10 DISPOSAL INSTRUCTION This product is subject to the WEEE disposal regulations and cannot be disposed of with regular waste. Please contact PEQLAB Biotechnologie GmbH or your local distributor for disposal of peqSTAR thermal cyclers. Waste electronic equipment is not useless waste. Environmentally friendly disposal can help to retrieve valuable resources. Thereby you contribute to the protection of the environment and human health. NOTES PEQLAB_v0612E 13 INHALTSVERZEICHNIS 15 15 16 16 16 16 16 17 17 17 18 18 21 22 23 24 25 25 25 25 26 26 PEQLAB_v0612D 14 1 2 LIZENZHINWEIS This instrument is licensed for research and development and for uses other than human in vitro diagnostics under one or more of the following patens of Applera Corporation: U.S. Patent Nos. 5,656,493, 5,038,852, 5,333,675, 5,475,610 (claims 1-159 and 164-166), 6,703,236 (claims 7-10) and 7,238,517 or corresponding claims in their non-U.S. counterparts. No right is conveyed expressly, by implication or by estoppels under any other patent of Applera, including but not limited to U.S. Patent No. 6,814,934 and its non-U.S. counterparts, which describe and claim thermal cyclers capable or real-time detection. PEQLAB_v0612D 15 3 4 4.1 PEQLAB_v0612D 16 Kurzanleitung peqSTAR Thermocycler 5 5.2 PEQLAB_v0612D 17 Kurzanleitung peqSTAR Thermocycler 5.3 Inkubationsmodus ('Run') 5.4 PEQLAB_v0612D Programmname Programm-Fenster 18 Loop Kombiniert Open Loop, Close Loop, Temperature, Ramp und Gradient. Open Lid Pause Heat Lid Store Temperature Open Loop Ramp Close Loop Heat Lid 03 07 Loop 04 Temp PEQLAB_v0612D 19 06 Temp 07 Loop-Funktion beenden mit 'Exit Loop'. Loop Open 04 05 06 07 Close Loop 08 Ramp 10 Store PEQLAB_v0612D 20 5.5 PEQLAB_v0612D 21 Clock User Accounts Hide/Show Curser Sounds Protocol (OFF/ON) Firmware Display Settings 5.6 PEQLAB_v0612D 22 Kurzanleitung peqSTAR Thermocycler 6 TECHNISCHER SERVICE UND BESTELLINFORMATIONEN Produkt peqSTAR 96 Universal peqSTAR 96 Universal Gradient peqSTAR 96 HPL peqSTAR 96 HPL Gradient peqSTAR 384 HPL peqSTAR in situ PEQLAB_v0612D 23 95-95002 95-96002 95-95002-HPL 95-96002-HPL 95-950384-HPL 95-950FLAT 7 TECHNISCHE DATEN PEQLAB_v0612D 24 Kurzanleitung peqSTAR Thermocycler 8 8.1 Kalibrierung PEQLAB_v0612D 25 Kurzanleitung peqSTAR Thermocycler 9 NOTIZEN PEQLAB_v0612D 26 D AT UK USA PEQLAB Biotechnologie GmbH, 91052 Erlangen, Freecall (D): 0800 100 20 16, [email protected], www.peqlab.de PEQLAB Biotechnologie GmbH, 6404 Polling, Tel: +43 (0) 5238 84 169, [email protected], www.peqlab.at PEQLAB Ltd., Southampton SO31 7ZN, Freecall (UK): 0808 20 21 302, [email protected], www.peqlab.co.uk PEQLAB LLC, Wilmington, DE 19810, Toll-Free (US): 877 737 5220, [email protected], www.peqlab.us Creating the future together.
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