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.
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.
