产品号 #76021_C
用于评估CRISPR-Cas9基因组编辑效率
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用于评估CRISPR-Cas9基因组编辑效率
Polymerase, buffers, and dNTPs for high-fidelity PCR amplification
DNase- and RNase-free water for molecular biology applications
For removing nucleases from DNA/RNA samples and terminating enzymatic reactions
For purification of genomic DNA from cells or tissue
For extraction of DNA from agarose gels, or clean-up of PCR products and restriction enzyme digestions
Compatible antibodies for purity assessment of isolated cells
ArciTect™ T7 核酸内切酶 I 是检测由 CRISPR-Cas9 产生的插入或缺失(INDELs)等基因组编辑的首选酶。ArciTect™ T7 核酸内切酶 I 试剂盒包含 ArciTect™ T7 核酸内切酶 I 和 ArciTect™ T7 核酸内切酶 I 缓冲液 (10X),经测试和验证,可与 ArciTect™ CRISPR-Cas9 基因组编辑系统配合使用。ArciTect™ T7 核酸内切酶 I 能识别并切割错配 DNA、十字形 DNA 结构、Holliday 结构或连接区、异源双链 DNA,以及低效切割的缺口双链 DNA。由于酶切效率与特定位点产生的 INDEL 数量成正比,因此 ArciTect™ T7 核酸内切酶 I 试剂盒可用于快速、经济地评估基因编辑效率。
细胞类型
多能干细胞
种属
人
应用
基因组编辑
研究领域
干细胞生物学
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.
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种属 | Human |
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Cas9 核酸酶,用于在 CRISPR-Cas9 基因组编辑中产生双链断裂
定制设计的 CRISPR RNA,用于 CRISPR-Cas9 基因组编辑中的guide RNA 生成
反式激活crRNA用于CRISPR-Cas9基因组编辑中的guide RNA生成
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