Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)
Type
Blog Post
...Detection of edited BRAF genomic loci in Cas9-expressing A549 cells transduced with BRAF gRNA at various ...To use this tool to detect a BRAF gene edit, simply PCR-amplify the BRAF locus from the edited cells (...product should contain the edited BRAF sequence), and PCR-amplify the BRAF locus from control cells (the ... the 428 bp band indicates edits in the BRAF gene in the BRAF-gRNA transduced cells, and no such edits...-targeted) BRAF sites (lanes 3, 4) were used as controls. Duplexes were generated from BRAF-edited and.../Cas9 gene edit. I wanted to inactivate the gene BRAF (a kinase implicated in several human cancers) in...editing, don’t expect a 100% knockout population.
BRAF gRNA delivery
Overview: In practice you would select...