What is the standard KASP thermal cycle program?

KASP™ chemistry utilises a two-step touchdown PCR method, with the elongation and annealing steps incorporated into a single step.  A 15 minute activation is required (94°C), followed by 10 cycles of touchdown PCR and 26 cycles of standard 2-step PCR. The standard KASP thermal cycle conditions are detailed in the table below.

Step Description Temperature Time Number of cycles per step
 1  Activation  94°C  15 min  1
 2  Denaturation  94°C  20 sec  10
 Annealing / Elongation  61 - 55°C  1 min (drop 0.6°C per cycle)
 3  Denaturation  94°C  20 sec  26
 Annealing / Elongation  55°C  60 sec


Please ensure that you check the cycling conditions in your assay information pack to ensure that your assay does not have any specific cycling conditions.  Some assays may require alternative cycling conditions due to high or low %GC content.