What should be the temperature for PCR machine lid during ligation overnight at 16c? Will the usual lid temperature at 105c affect the ligase performance?
keeping the lid at 5c above the highest programmed temperature should be enough to stop evaporation and condensation of the sample
The temperature of the PCR machine lid during ligation overnight at 16°C should ideally be maintained at a temperature that prevents condensation from forming on the lid, as condensation can potentially affect the reaction. This typically means setting the lid temperature slightly above the reaction temperature, but not high enough to cause evaporation or denaturation of reaction components.
For overnight ligation at 16°C, you might set the lid temperature around 17-18°C to prevent condensation without significantly affecting the reaction. However, it's essential to verify with the specific PCR machine model you're using, as lid temperatures can vary between instruments.
Most PCR machines do not allow a temp lower than 40. For overnight ligation at 16 I just turn off the Led heating and it has worked out fine.
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