Cas9 Nuclease NLS, S. pyogenes, is an RNA-guided endonuclease that catalyzes site-specific cleavage of double stranded DNA. The location of the break is within the target sequence 3 bases from the NGG PAM (Protospacer Adjacent Motif). The design of single guide RNA (sgRNA) is dependent on the target region close to the PAM site. Even if you pick a target sequence that fulfills all of the described requirements, sgRNA specificity and activity is unpredictable. Therefore, it is often recommended that multiple, different sgRNAs be designed to target a gene of interest. The sgRNA Screening Kit provides a simple, reliable, and rapid method for assessing sgRNA efficiency before cell transduction, allowing you to identify the highly effective CRISPR sgRNA.Applications:Evaluate the efficiency of sgRNAs for CRISPR/Cas9 in vitro studies and identify the highly effective sgRNAs before cell transfection.Kit Components:Cas9 Nuclease: 30rxn:15ul, 100rxn: 50ul10X Reaction Buffer: 30rxn:100ul, 100rxn: 300ulPositive Control sgRNA: 30rxn: 3ug, 100rxn: 10ugPositive Control Substrate: 30rxn: 60ug, 100rxn: 200ulRNase-free H2O: 30rxn: 1ml, 100rxn: 1mlStorage and Stability:Store at -20°C upon receipt. positive control sgRNA should be aliquoted and stored at -70°C, to avoid repeated freeze/thaw cycles.