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 GenCrispr 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.
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Research purposes only.
| Storage |
1. The positive control sgRNA is provided as a freeze-dried powder that should be stored at -80°C once received. It is recommended to be used within 3 months after receiving. Redissolve with RNase-free water before using. 2. The activity of the positive control sgRNA will decrease after free-thaw cycles. Please aliquot and store at-80°C, to avoid repeated freeze/thaw cycles. 3. There commended storage temperature of GenCrispr Cas9 Nuclease and 10X Reaction buffer is -20°C. |
| Evaluate the efficiency of sgRNAs for CRISPR/Cas9 in vitro studies and identify the highly effective sgRNAs before cell transfection. |
gRNA efficiency screening using GenCRISPR sgRNA screening kit.
Twelve unique synthesized gRNAs were incubated with Cas9 nuclease for 1 h at 37°C according to the instructions in theGenCRISPR sgRNA screening kit manual. Upon separating the reactions by agarose gel electrophoresis, only gRNA9 was observed to induce cleavage.
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