NEB does not offer sgRNA synthesis services or plasmids for sgRNA cloning. For cloning, we recommend visiting Addgene to obtain sgRNA plasmids. NEB tools enable a broad variety of methods for manipulating these plasmids. To learn more about methods for rapidly swapping out targeting sequences in plasmids please visit this page: https://www.neb.com/applications/genome-editing/sgrna-template-construction-for-cas9-gene-editings.

For quickly transcribing sgRNAs without the need for cloning or PCR, we recommend the EnGen sgRNA Synthesis Kit, S. pyogenes (NEB #E3322).

Alternatively, oligonucleotide templates encoding sgRNA transcribed by a T7 promoter can be used in conjunction with an in vitro transcription kit such as the NEB’s HiScribe^® T7 Quick High Yield RNA Synthesis Kit (NEB #E2050) to quickly and efficiently produce guide RNAs.