- 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.