denature 94c for 2 minute then 10 cycle for 94c 30sec and annealing temp 71c to 61c then 15 cycle 61c annealing 72 c extension and final extension 72c for 4minute
Hi Khalid Akhtar
Touchdown is achieved by setting the ramp rate on the system. You can create a regular program with all the steps you have mentioned, only keeping temperature of the touchdown step as 71C. Then for that specific step, select ramp rate to be -1C/ cycle.
Hope it helps,
Subham.
To make a touchdown PCR profile on a thermal cycler, you can follow these steps:
It's important to note that the specific parameters of the touchdown PCR profile may need to be adjusted depending on the specific PCR application, so it's always a good idea to consult the literature or a PCR expert to optimize the protocol for your particular experiment. Additionally, it's important to properly calibrate your thermal cycler to ensure accurate temperature control during the PCR reaction.
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