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.
Product table
| PRODUCT | Prod. # | SIZE | INFO | |
|---|---|---|---|---|
| EnGen® sgRNA Synthesis Kit S. pyogenes | E3322S | 20 rxns |  |
 |
| E3322V | 10 rxns | |
|
|
| HiScribe™ T7 High Yield RNA Synthesis Kit | E2040S | 50 rxns | |
|
| HiScribe™ T7 Quick High Yield RNA Synthesis Kit | E2050S | 50 rxns | |
|
| Q5® High-Fidelity DNA Polymerase | M0491S | 100 units | |
|
| M0491L | 500 units | |
|
|
| Q5® Hot Start High-Fidelity DNA Polymerase | M0493S | 100 units | |
|
| M0493L | 500 units | |
|
|
As of: 01.01.2024
Further information can be found in our Technical Resources section or at neb.com. Information on trademarks can be found here.
