FAQ: Why do I observe incomplete binding with SNAP dCas9 NLS, S. pyogenes?

Incomplete binding may be due to the following factors:
  • Incorrect ratio of dCas9 to guide RNA, and target site-For complete binding we recommend a 10:10:1 or higher molar ratio (SNAP dCas9 NLS : guide RNA : target site).
  • Suboptimal guide RNA sequence-Verify the sequence and design of the guide RNA.
  • Poor quality guide RNA-Verify the integrity of the guide RNA by gel electrophoresis.
  • Buffer incompatibility-Please use the 10X NEBuffer 3.1 included with the enzyme.