FAQ: What are the main causes of blunting reaction failure?

The following conditions can cause exonuclease activity to overwhelm the polymerase activity, causing recessed instead of blunt ends:

  • Failure to add nucleotides, or the nucleotide concentration is too low
  • Adding too much enzyme
  • Incubating for too long
  • Incubating at temperatures greater than the recommended temperature
  • Heat inactivating without EDTA

Please see the recommended blunting protocol for 

 
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