CRISPRcraft™ S.p. Cas9 Nuclease Control Kit
Human HPRT positive controls for CRISPRcraft™ S.p. Cas9 Nuclease
- Take control of your CRISPR experiments with one kit containing controls for both in vivo and in vitro Cas9 experiments
- In vivo assays: kit includes positive control guide RNA and PCR primers for post-editing target site amplification
- In vitro assays: kit includes positive control DNA substrate, guide RNA and reaction buffer
- Kits do not include Cas9 nuclease enzyme
- Looking for CRISPRcraft™ S.p. Cas9 Nuclease?
CRISPRcraft™ S.p. Cas9 Nuclease Control Kit contains convenient controls for both in vivo gene editing experiments, and in vitro Cas9 cleavage assays.
|Kit Component(s) ||Use with In Vitro Assays ||Use with In Vivo Assays |
|Cas9 Control HPRT Guide cRNA and Universal tracrRNA || || |
|10X RGEN Buffer || || |
|Control HPRT Substrate DNA || || |
|Human HPRT Forward / Reverse PCR Primers || || |
The kit includes control guide RNA for human HPRT, supplied as dual guide crRNA and tracrRNA components that are quickly annealed before use. Combine the duplexed guide with CRISPRcraft S.p. Cas9 Nuclease (sold separately) to form RNP complexes, then use the HPRT-targeting RNP complexes as positive controls in gene editing experiments in human cells. We’ve also included forward and reverse PCR primers for easy post-editing amplification of the human HPRT locus. The resulting HPRT PCR amplicon can be used for mutation detection assays, such as the T7 Endonuclease 1 assay.
The RNP complexes formed with CRISPRcraft S.p. Cas9 and control HPRT guide RNA can also be used in conjunction with the supplied Control HPRT Substrate DNA and 10X RGEN Buffer for in vitro Cas9 cleavage assays. Expected results of the in vitro cleavage assay are shown in Figure 1.
CRISPRcraft S.p. Cas9 Nuclease In Vitro Titration Assay
Figure 1: In Vitro Assay. In Vitro Assay. A dilution series of CRISPRcraft S.p. Cas9 Nuclease was used to generate RNP complexes with Cas9 Control Human HPRT cRNA and Cas9 Control Universal tracrRNA. RNP complexes were incubated in triplicate with Control HPRT Substrate DNA for 10 minutes at 37°C in 1X RGEN Buffer. Results were visualized by agarose gel electrophoresis (A) and % DNA cleavage was calculated from band intensity (B).
Kit contains Cas9 Control HPRT Guide crRNA and Cas9 Universal tracrRNA, Human HPRT Forward and Reverse PCR Primers, Control HPRT Substrate DNA and 10X RGEN Buffer. The Kit does not contain Cas9 enzyme.