Reaction Conditions

1X Cas9 Nuclease Reaction Buffer
Incubate at 37°C

1X Cas9 Nuclease Reaction Buffer
20 mM HEPES
100 mM NaCl
5 mM MgCl₂
0.1 mM EDTA
(pH 6.5 @ 25°C)

Storage Temperature

-20°C

Storage Conditions

300 mM NaCl
10 mM Tris-HCl
0.1 mM EDTA
1 mM DTT
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

65°C for 5 min

Companion Products

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

1. 1,000 nM is equal to 159 ng/μl.

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

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?

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 (NEB #M0386)

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

• NEBioCalculator

• Construction of an sgRNA Cas9 expression vector

• 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

• 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

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

• Cas9 Nuclease, S. pyogenes
• Cas9 Nuclease Reaction Buffer

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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.