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T7 Endonuclease I (T7EI) is a structure-selective enzyme that recognizes and cleaves mismatched DNA, cruciform DNA structures, Holliday structures or junctions, heteroduplex DNA, as well as nicked double-stranded DNA. This enzyme has a preference for single stranded over double stranded DNA with cleavage occurring at the first,second or third phosphodiester bond that is 5′ to the mismatch. In CRISPR/TALEN experiments, T7E1 can accurately recognize insertions and deletions of ≥2 bases that are generated by non-homologous end joining (NHEJ) activity. GeneCopoeia’s T7 endonuclease I assay kits, which contain T7E1 and positive controls for both target PCR and indel mismatch cleavage, provide a simple and fast workflow for CRISPR/TALEN validation and screening. T7E1 is compatible with a broad range of PCR buffers and does not usually require purification of the PCR product prior to digestion. By incubating re-annealed PCR fragments with T7 endonuclease I, the presence of two shorter bands of predicted sizes usually means that CRISPR/TALEN has successfully introduced indel mutations at the targeted chromosomal site. T7 endonuclease I assay kits can also be used with our Target site PCR kits and Target site PCR cloning kits for IndelCheck™ CRISPR/TALEN insertion or deletion (indel) detection system. Advantages
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T7 Endonuclease I Assay Kit
Introduction
