Hi, have a look at this publication:
Title: Touchdown PCR for increased specificity and sensitivity in PCR amplification
Authors: Korbie DJ, Mattick JS
Nat Protoc. 2008;3(9):1452-6
Cheers, Nadine
Touch down PCR is used to avoid non-specific amplification. In this annealing temperature is gradually reduced (1-2°C /every second cycle). The initial annealing temperature should be several degrees above the Tm of the primer. The annealing temperature is then gradually decreased until it reaches the calculated annealing temperature of the primer.
@Himanshu Aggarwal
You mean to say that Touchdown PCR is an alternative for Gradient PCR ?
In a touch down PCR you gradually reduce your annealing temperature. You start your PCR at slightly higher temperature say 62 deg C. You allow this for a few cycles say 3 cycles and from fourth cycle onwards you reduce your annealing temperature to 60 deg C. After next set of 3 cycles you again reduce you annealing to 58 deg C and it goes on like this. Its mainly carried out to reduce amplification of non specific bands. Hope the explanation helps........Good luck.
Hi, have a look at this publication:
Title: Touchdown PCR for increased specificity and sensitivity in PCR amplification
Authors: Korbie DJ, Mattick JS
Nat Protoc. 2008;3(9):1452-6
Cheers, Nadine
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