Cas9 Nuclease, 300 pmoles

Features
The Cas9 protein is a recombinant endonuclease from Streptococcus pyogenes. 

Basic description

The Cas9 protein is a recombinant endonuclease from Streptococcus pyogenes with a molecular weight of 150 kDa. Cas9 endonuclease in complex with RNA guides (crRNA duplex: tracrRNA) or single sgRNA catalyzes site-specific hydrolysis of the phosphodiester bond in double-stranded DNA (Figure). The cleavage is on the DNA chain between the third and fourth nucleotides from the PAM sequence (NGG is a motif adjacent to the protospacer) with the formation of blunt ends. Recombinant Cas9 endonuclease does not contain a nuclear localization sequence.

Figure: DNA digestion with sgRNA:Cas9 complex.

Source: Cas9 endonuclease purified from E. coli strain containing a plasmid with the cloned full length gene of Cas9 Streptococcus pyogenes.

Quality control. Each batch of the enzyme is tested for enzyme activity, electrophoretic purity, and the absence of non-specific endonuclease activity. The purity of the enzyme is ≥95% (SDS-PAGE), nonspecific endonuclease activity is absent.

Storage buffer: 300 мМ NaCl, 50 мМ Tris-HCl, 0,1 мМ EDTA, 5 мМ β-mercaptoethanol, 50% glycerol (pH=7.5 @ 25°C).

Standard buffer for the plasmid DNA hydrolysis reaction: 20 mM HEPES, 125 mM KCl, 1 mM EDTA, 1 mM dithiothreitol, 6 mM MgCl2, 7% glycerol (pH 7.5 @ 25 ° C).

The optimum reaction temperature: 37°C.

Enzyme inactivation: 65°C for 5 minutes.

Standard product concentration: 20 μM (20 pmol/μl)

Minimum packing unit: 300 pmoles (15 μl)

Areas of use

  • Genomic editing, CRISPR / Cas9 technology

Benefits of using

Storage

Transportation

Storage and transportation at -20ºС.

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