Guide-it Genotype Confirmation Kit
- Distinguish monoellelic and biallelic CRISPR/Cas9 induced mutants
- Quickly identify the clone with the right genotype
- In vitro cleavage method using recombinant Cas9 nuclease
Identify monoallelic and biallelic mutants after CRISPR/Cas9 gene editing
The CRISPR/Cas9 system can be used to generate insertions or deletions (indels) that can result in gene knockouts. In most cells, there are two copies of any given gene, and indels can be generated in either one or both alleles. The Guide-it Genotype Confirmation Kit is used after gene editing to determine if a given clone has mutations in one allele (monoallelic), both alleles (biallelic), or is unchanged (wild type).
Using this kit after gene editing speeds up the screening a large number of clones to find those with the desired genotype.
How does it work?
First you amplify the target site from the cells edited with Cas9 and sgRNA. The resulting PCR fragment is then used in an in vitro cleavage reaction with recombinant Cas9 and the same sgRNA. The genotype can be determined by resolving the cleavage products on an agarose gel.