sgRNA Template Construction for Cas9 Gene EditingReturn to Genome Editing
Along with Cas9 nuclease, CRISPR experiments require the introduction of an sgRNA containing an approximately 20-base sequence specific to the target DNA 5′ of a non-variable scaffold sequence. sgRNA can be delivered as RNA or by transforming with a plasmid with the sgRNA-coding sequence under a promoter. A number of strategies have been developed to quickly swap out the 20 base sequences allowing convenient sgRNA cloning using NEB products.
Construction of an sgRNA-Cas9 Expression Vector via an ssOligo Bridge
Learn how you can use NEBuilder HiFi to generate an sgRNA-Cas9 expression vector with a single-stranded oligo bridge.