Uracil-N-glycosylase (UNG) has activity in the temperature range of 4 degrees C to 55 degrees C, and is not completely inactivated by incubation at 95 degrees C. It is recommended that the annealing temperature be at or above 55 degrees C when using this enzyme with any of our PCR or Sequence Detection Systems kits to avoid possible degradation of newly synthesized dU-containing PCR products by residual UNG activity. For this reason, it is also recommended that the post-PCR soak be maintained at 72 degrees C (instead of 4 degrees C) if analysis of the PCR product will be performed using a gel-based method.
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PCR and cDNA Synthesis Support Center.