Cas9 Nuclease, S. pyogenes

Companion Products

• EnGen® Mutation Detection Kit
• EnGen® sgRNA Synthesis Kit, S. pyogenes

1. 1,000 nM is equal to 159 ng/μl.
2. If planning to use higher concentration Cas9 Nuclease, S. pyogenes (M0386T and M0386M) for in vitro digestion of DNA, the enzyme should be diluted to 1 µM using Diluent B (NEB #B8002S) prior to the reaction assembly.

1. Jinek M. et al. (2012). Science. 816-821 doi: 10.1126/Science.1225829. Epub 2012 Jun 28.. PubMedID: 22745249

1. In vitro digestion of DNA with Cas9 Nuclease, S. pyogenes (M0386)
2. Using recombinant Cas9 nuclease to assess locus modification in genome editing experiments (#M0386)

• Bridging dsDNA with a ssDNA Oligo and NEBuilder® HiFi DNA Assembly to create an sgRNA-Cas9 Expression Vector

1. Why do I observe incomplete digestion?
2. Why does digestion efficiency differ between two SgRNAs?
3. Does NEB provide plasmids for gRNA cloning?
4. Does Cas9 Nuclease contain a nuclear localization signal (NLS)? 
5. Is Cas9 Nuclease fused with a N-terminal 6XHis tag?
6. How do I dilute the enzyme to 1 μM for in vitro reactions?

• Haussmann IU, Ustaoglu P, Brauer U, Hemani Y, Dix TC, Soller M. (2018) Plasmid-based gap-repair recombineered transgenes reveal a central role for introns in mutually exclusive alternative splicing in Down Syndrome Cell Adhesion Molecule exon 4 Nucleic Acids Res; Dec 12, PubMedID: 30541104 , DOI: 10.1093/nar/gky1254
• Lee, NC., Larionov, V., Kouprina, N. (2015) Highly efficient CRISPR/Cas9-mediated TAR cloning of genes and chromosomal loci from complex genomes in yeast Nucleic Acids Res; 43(8), e55. PubMedID: 25690893

• Pardee P, Green A A, Takahashi M K, Braff D, Lambert G, Lee J W, Ferrante T, Ma D, Donghia N, Fan M, Daringer N M, Bosch I, Dudley D M, O’Connor D H, Gehrke H, Collins J J (2016) . Rapid, Low-Cost Detection of Zika Virus Using Programmable Biomolecular Components Cell.; DOI: 10.1016/j.cell.2016.04.059

• CRISPR/Cas9 & Targeted Genome Editing: New Era in Molecular Biology

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

• M0386S_L_v1
• M0386T_M_v1

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

• M0386M_v1_10011915
• M0386T_v1_10011917
• M0386S_v1_10018044
• M0386M_v1_10028145
• M0386T_v1_10028147
• M0386S_v1_10035061
• M0386M_v1_10043718
• M0386T_v1_10043721
• M0386M_v1_10057223
• M0386T_v1_10057222
• M0386S_v1_10057218
• M0386M_v1_10062631
• M0386T_v1_10062630

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• Cas9 Nuclease, S. pyogenes
• NEBuffer™ 3.1

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.