Genomic Deletions in Mammalian Cell Lines
Type
Collection
...example, sgRNA-A’s protospacer sequence and PAM happen to fall on the bottom (Crick) strand while sgRNA-B’s...primers flanking each sgRNA target site (within 150 - 350 bp) to amplify the sgRNA target site to examine...complement: 1.0 μl sgRNA 24- or 25-mer oligo (100 μM; see step 1.4 ), 1.0 μl sgRNA 24- or 25-mer reverse...composed of a polyclonal population exposed to sgRNA-A and sgRNA-B (see step 5 ). The purpose of the following...delivery of a pair of chimeric single guide RNAs (sgRNAs) to create two double strand breaks (DSBs) at a...posted at the top of the page. CRISPR Design Design sgRNAs manually or using freely available online tools...non-coding elements. For creating a gene knockout, two sgRNA located within exons will enrich even monoallelic...