FAQ: Why am I observing limited or no digestion with EnGen® Lba Cas12a (Cpf1)?

Incomplete digestion may be due to the following factors:

  • Incorrect ratio of EnGen Lba Cas12a (Cpf1) to gRNA and/or target site. For complete digestion we recommend a 10:10:1 or higher molar ratio of EnGen Lba Cas12a (Cpf1) : gRNA : target site.
  • Suboptimal sequence of the gRNA. Verify the sequence and design of the gRNA.
  • Poor quality gRNA. Verify the integrity of the gRNA by gel electrophoresis.
  • Use of a suboptimal buffer. Please use EnGen Lba Cas12a Diluent, which is included with the enzyme.
  • When diluting 100 µM EnGen Lba Cas12a (Cpf1) for use in in vitro cleavage assays, supplementing reactions with 10 mM DTT can in some cases improve substrate cleavage.