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.
Plasmids containing sgRNA sequences can be constructed using a variety of methods. Common to each is the requirement to introduce an approximately 20 base target sequence downstream of a promoter. Guide RNA templates can then be used as templates for in vitro transcription or directly introduced.
As of: 03.01.2022
* Permanent low prices – no further discounts apply. Offer closes 31.12.2022.