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Thermal Cycler FAQs
Q: Why is a temperature gradient required?
A: The annealing temperature of the specific primers used in DNA amplification often requires optimisation. Instead of running multiple experiments with different annealing temperatures and different MgCl2 and primer concentrations the gradient thermal cycler allows the testing of 12 different annealing temperatures (8 different reaction conditions) simultaneously.
Q: What is "Hot start"?
A: The Hot Start step is used to pause the machine at a specific temperature, typically around 70°C, after the initial template denaturation. The reason is to allow the manual addition of unmodified Taq DNA polymerase which may loose activity if added during the initial 5min denaturation. Heat-activated Taq or Hot Start enzymes do not require this step.
Q: How is it possible to connect a single thermal cycler to a computer?
A: If only one cycler is going to be connected then an RS232 cable (part code FGEN232) is required. For the TC-3000X and TC-3000G cyclers a USB cable is required (part code 6205046)
Q: How is it possible to connect multiple thermal cyclers to a computer?
A: The table below indicates which power supply, (when there is no TC-512 in the chain), cables and convertor are required for various set-ups. If further assistance is required please contact the Technical Support team at technehelp@bibby-scientific.com
| TC Model | Additional Units | No. Extra Units | Power Supply | Cable Set | Convertor | Additional cables |
|---|---|---|---|---|---|---|
| TC-3000, TC-312 or TC-412 | TC-3000, TC-312 or TC-412 | 1 | FGEN485D | FGENTWO | FTGEN485 | Not required |
| TC-3000, TC-312 or TC-412 | TC-3000, TC-312 or TC-412 | 3 | FGEN485D | FGENFOUR | FTGEN485 | Not required |
| TC-3000, TC-312 or TC-412 | TC-3000, TC-312 or TC-412 | 9 | FGEN485D | FGENTEN | FTGEN485 | Not required |
| TC-3000, TC-312 or TC-412 | TC-3000, TC-312 or TC-412 | 10 | FGEN485D | FGENTEN | FTGEN485 | FGENONE |
| TC-512 | TC-512 | 1 | Not required | 6103557 | FTGEN485 | Not required |
| TC-512 | TC-512 | 2 | Not required | 6103557 | FTGEN485 | 6103557 |
| TC-512 | TC-3000, TC-312 or TC-412 | 1 | Not required | FGENONE | FTGEN485 | Not required |
| TC-512 | TC-3000, TC-312 or TC-412 | 2 | Not required | FGENTWO | FTGEN485 | Not required |
| TC-512 | TC-3000, TC-312 or TC-412 | 3 | Not required | FGENTWO | FTGEN485 | FGENONE |
Q: What is the incremental function?
A: Incremental timing and temperature are used to increase or decrease either the time or temperature incrementally over the number of cycles in a stage. Incrementation of extension time is used with "Long PCR" which is when large template fragments are to be amplified (e.g. 27kb lambda DNA, 40kb genomic DNA). Decremental temperature is used for protocols such as "Touchdown PCR" where the first cycle starts with a high annealing temperature and over the number of cycles there is a gradual decrease in the temperature. This ensures that only the specific product is amplified.
Q: What is a gradient thermal cycler?
A: Gradient blocks enable a particular temperature step in a protocol to be altered so that the temperature varies across the block. By specifying a temperature and the gradient to be applied, each column will achieve a different temperature, with the set temperature in the middle block, the lowest on the left and the highest on the right. The gradient is the total difference across the block, for example if the set temperature was 60°C and the gradient 10°C then the temperatures would range from 55°C to 65°C from left to right.
Q: What is the 'Pause' function at the start of the program used for?
A: Some users prefer to preheat the heated lid before placing the samples into the unit. The pause feature is used to stop the unit after the 4 minute heated lid preheat step. It will also sound an audible alarm indicating that the machine is ready for the sample tubes or plate to be added. Pressing the "Pause" key followed by the "Enter" key will commence the remaining program.
TC-Plus FAQ's
Q: How do I adjust the pressure of the heated lid for my tubes or plates?
A: No adjustment is necessary. The lid automatically applies the correct pressure. Ensure that all consumables used in the block are of the same height.
Q: Do I need a different block for fully skirted plates?
A: No. The TC-PLUS 96-well block will accommodate fully skirted as well as half-skirted and non-skirted PCR plates.
Q: What is `Sample Cooling'?
A: Sample cooling can be applied during the lid pre-heat phase. This maintains the block at 4°C while the lid is heating to temperature and avoids any increase in sample temperature due to the heated lid before the thermal cycling program begins.
Q: How does TERS™ work?
A: TERS™ harnesses the released heat during the cooling phase for use in the next heating phase. By switching the fans off for a very short time at the end of the cooling period the heat is not removed from the heat sink, but is instead stored as heat energy. Heat is therefore available as soon as the Peltiers switch from cooling to heating. This results in faster heating, requires less energy to heat the block and subsequently costs less to run.
Q: What is `Hot start'?
A: The Hot Start step is used to pause the machine at a specific temperature, typically around 70°C, after the initial template denaturation. The reason is to allow the manual addition of unmodified Taq DNA polymerase which may loose activity if added during the initial denaturation step. Heat-activated Taq or Hot Start enzymes do not require this step.
Q: What is the incremented time and temperature function for?
A: Incremented/decremented time and temperature are used to increase or decrease either the time or temperature incrementally over the number of cycles in a stage. Incrementation of extension time is used with `Long PC' which is when large template fragments are to be amplified (e.g. 27kb lambda DNA, 40kb genomic DNA). Decremented temperature is used for protocols such as 'Touchdown PCR' where the first cycle starts with a high annealing temperature and over the number of cycles there
is a gradual decrease in the temperature. This ensures that only the specific product is amplified.
Q: What is a gradient thermal cycler?
A: Gradient blocks enable a particular temperature step in a protocol to be altered so that the temperature varies across the block. By specifying a temperature and the gradient to be applied, each column will achieve a different temperature, with the set temperature in the middle of the block, the lowest on the left and the highest on the right. The gradient is the total difference across the block, for example if the set temperature was 60°C and the gradient 10°C then the temperatures would range from approximately 55°C to 65°C from left to right.
Q: Why is a temperature gradient required?
A: The annealing temperature of the specific primers used in DNA amplification often requires optimisation. Instead of running multiple experiments with different annealing temperatures the gradient thermal cycler allows the testing of up to 12 different annealing temperatures simultaneously.
