In vitro DNA cleavage assay with GenCrispr NLS-Cas9-D10A Nickase
Reactions were set up according to recommended conditions, and cleavage products were resolved on a 1% agarose gel. Input DNA is pUC57 plasmid DNA (OC, open circular; LIN, linearized; SC, supercoiled)
GenCrispr NLS-Cas9-D10A Nickase
GenCrispr NLS-Cas9-D10A Nickase is a mutation form of Cas9 Nuclease. Cas9 nuclease is an RNA-guided endonuclease that can catalyze cleavage of double stranded DNA. This kind of targeted nuclease is a powerful tool for genome editing with high precision. Cas9 nuclease cleaves the double strand DNA generating two break sites based on its two active domains. NLS-Cas9-D10A Nickase is a mutation form of Cas9 Nuclease which makes one active domain deactivated, thus it can only cut one single strand DNA that is complementary to the guide-RNA, producing one single strand cut. Combined with two different gRNA, NLS-Cas9-D10A Nickase produces two cut sites respectively and causes a double strand break. Compared with the wild type Cas9 nuclease, the two-gRNA guided cleavage can significantly reduce the off target effects. Product Source: GenCripsr NLS-Cas9-D10A Nickase is highly purified mutant proteins expressed in an E. coli strain carrying a plasmid encoding the mutated Cas9 gene from Streptococcus pyogenes. Key features: DNA-free: no external DNA added to system. High cleavage efficiency: NLS ensures the efficient entry of Cas9 protein into nuclei. Low off target: Double gRNA-guided cleavage and transient expression of Cas9 nuclease. Time-saving: no need for transcription and translation. With this Cas9 Nuclease, you can: Screening the highly efficient and specific targeting gRNAs using in vitro DNA cleavage. In vivo gene editing combined with specific gRNA by electroporation or injection.
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