ArciTect™ T7 Endonuclease I Kit_产品中心_北京砺达辰生物科技有限公司

分子生物学

ArciTect™ T7 Endonuclease I Kit

For estimation of CRISPR-Cas9 genome editing efficiency

概要

技术资料

数据及文献

概要

ArciTect™ T7 Endonuclease I is the preferred enzyme for detecting genome editing such as insertions or deletions (INDELs) generated by CRISPR-Cas9. ArciTect™ T7 Endonuclease I Kit is comprised of ArciTect™ T7 Endonuclease I and ArciTect™ T7 Endonuclease I Buffer (10X), which have been tested and validated for use with the ArciTect™ CRISPR-Cas9 genome editing system. ArciTect™ T7 Endonuclease I recognizes and cleaves mismatched DNA, cruciform DNA structures, Holliday structures or junctions, heteroduplex DNA, and, less efficiently, nicked double-stranded DNA. Since the cleavage efficiency is proportional to the number of INDELs created at a specific DNA target, ArciTect™ T7 Endonuclease I Kit is used to estimate gene-editing efficiency in a rapid and cost-effective manner.

技术资料

Document Type	产品名称	Catalog #	Lot #	语言
Product Information Sheet	ArciTect™ T7 Endonuclease I Kit	76021, 76022	All	English
Safety Data Sheet 1	ArciTect™ T7 Endonuclease I Kit	76021, 76022	All	English
Safety Data Sheet 2	ArciTect™ T7 Endonuclease I Kit	76021, 76022	All	English

数据及文献

Data

Figure 1. INDEL Detection by T7 Endonuclease I Assay

Human embryonic stem (ES) and induced pluripotent stem (iPS) cells (A) and T cells (B) were edited using ArciTect™ Cas9 Nuclease (Catalog #76002) and ArciTect™ Human HPRT Positive Control Kit (Catalog #76013), and INDEL formation was assessed using ArciTect™ T7 Endonuclease I Kit. Following CRISPR-mediated editing at the HPRT locus, genomic DNA was isolated and a 1 kb region surrounding the target site was amplified by PCR using ArciTect™ Human HPRT Primer Mix (included with Catalog #76013). PCR products were purified, then denatured, re annealed, and cut with ArciTect™ T7 Endonuclease I. Samples were resolved on a 1% agarose gel, and band intensities were determined using a ChemiDoc™ MP Imaging System (Bio-Rad). Relative intensities of the full length (1083 base pairs [bp]) and T7 cleavage product bands (827 and 256 bp) were used to calculate the cleavage efficiency (%). Control: Uncut PCR product (no T7 added); Test, Donor 1, and Donor 2: T7-digested PCR product.