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Encompassing Amino Acids: 2-1368(end)
Application: Cas9 invitro cleavage assays to identify sgRNA with the highest cutting efficiency.
Assay Conditions: The Cas9 nuclease was incubated with sgRNA at 37°C for 10 minutes to form a functional RNP complex. A purified PCR product was then added at 37°C for 60 minutes, before Cas9 activity was visualized by agarose gel.
Description: Streptococcus pyogenes CRISPR associated protein 9 (Cas9), also known as SpCas9, GenBank Accession No. NC_002737.2:854751-858857, a.a. 2-1368(end) with N-terminal HIs-tag, expressed in an E. coli cell expression system. MW = 160 kDa.
Format: Aqueous buffer solution
Formulation: 20 mM HEPES, pH 7.5, 500 mM KCl, 20% Glycerol, 3 mM DTT.
Genbank: NC_002737.2:854751-858857
Purity: ≥90%
Storage Stability: At least 6 months at -80°C.
Supplied As: Aqueous buffer solution.
Tags: N-terminal His-tag
Uniprot: Q99ZW2
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Liang, X. et al., J Biotechnol. 2017; 241:136-146
2. Kim, S., et al., Genome Res. 2014; 24(6): 1012-1019