| Thermal Cyclers |
| How does one clean the Thermal Cycler block? |
| The heat/cool block, including wells and flat surfaces, should be cleaned regularly to ensure optimum heat transfer to the samples. Always clean the block if there has been a spillage. Use a cloth or cotton buds dipped in a fresh 50:50 water/isopropanol solution and make sure that no deposits are left in the wells. |
| Where should I place the tubes if I don't intend to use all the wells? |
| You can place the tubes in any location on the block. To ensure even pressure of the heated lid on your samples, it is necessary to place empty tubes at the corners of the reaction block. |
| How do I eliminate evaporation? |
| Evaporation, or sample loss, is caused by inadequate sealing. If you are using tubes, ensure they are sealed tightly or switch to a brand that has better sealing abilities. Sealing film on 96-well plates has the highest probability of evaporation, as film is more difficult to seal correctly. Using an applicator or roller to go over the film once it has been applied to the plate will ensure better adhesion. Also the use of a compression pad may be used to aid in the sealing when pressure is applied from the lid. |
| How do I eliminate condensation? |
| There are two types of condensation that can form in the tube. Condensation on the walls of the tube close to the block is common and won't affect your results. Condensation on the lid of the tubes is more of a concern and can be caused by inadequate lid pressure, inadequate lid contact or inadequate lid temperature. Check to ensure you have pressure on the tubes or switch to a flattop PCR tube. Lid temperature is recommended to be approximately 10-15°C higher than the block temperature (approximately 110°C) |
| What recommendations do you have to extend the life of the thermal cycler instrument? |
| The heating and cooling required for PCR reactions can put a lot of strain on the block. A misconception in the research world is that after the PCR reaction is complete, a final hold temperature of 4°C is required to deactivate the enzyme. A 10°C to 12°C final hold temperature is perfectly acceptable for many PCR applications and in some cases even room temperature will not affect the results. Minimizing or preventing extended holds (overnight and over a weekend) is another important factor to increase the longevity of the block. |
| Can poor yield be caused by the thermal cycler? |
| Generally, the thermal cycler will not affect your yield. The first thing to review is your sample preparation. Check that your reagents have not expired, pipetting volumes are accurate and you do not have any contamination. If you are assured that it is not your sample preparation, contact the manufacturer to help identify the issue. |
| What should I check over when I receive my new thermal cycler? |
| Always review the manual to ensure you understand how the instrument needs to be unpacked and set-up. |
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