FAQ: How can I synthesize a sequence-specific sgRNA for experiments using EnGen® Sau Cas9?

sgRNAs to be used with EnGen Sau Cas9 can be synthesized in vitro using the HiScribe™ T7 High Yield RNA Synthesis Kit (NEB #E2040) or the HiScribe T7 Quick High Yield RNA Synthesis Kit (NEB #E2050).  Double-stranded DNA templates can be either PCR products, synthetic double-strand DNA templates or linearized plasmids.

More detailed design recommendations can be found in the protocols section.

Alternatively, a synthetic sgRNA could be ordered from an RNA synthesis company.