ArciTect™ T7 核酸内切酶 I 试剂盒

用于评估CRISPR-Cas9基因组编辑效率

率先评论此产品

产品号 #（选择产品）

产品号 #76021_C

用于评估CRISPR-Cas9基因组编辑效率

产品组分包括

ArciTect™ T7 核酸内切酶 I 试剂盒，25 Reactions（产品号 #76021）

ArciTect™ T7 核酸内切酶 I，25 Reactions
ArciTect™ T7 核酸内切酶 I 缓冲液 (10X)，1 mL

ArciTect™ T7 核酸内切酶 I 试剂盒，125 Reactions（产品号 #76022）

ArciTect™ T7 核酸内切酶 I，125 Reactions
ArciTect™ T7 核酸内切酶 I 缓冲液 (10X)，1 mL

跳到结尾的图片库

跳转到图像库的开头

要查看实验方案所需的所有配套产品，请参阅《实验方案与技术文档》。

ArciTect™ High-Fidelity DNA Polymerase Kit
Polymerase, buffers, and dNTPs for high-fidelity PCR amplification
Nuclease-Free Water
DNase- and RNase-free water for molecular biology applications
Proteinase K Solution
For removing nucleases from DNA/RNA samples and terminating enzymatic reactions
Genomic DNA Purification Kit
For purification of genomic DNA from cells or tissue
Gel and PCR Clean-up Kit
For extraction of DNA from agarose gels, or clean-up of PCR products and restriction enzyme digestions
Labeling Antibodies
Compatible antibodies for purity assessment of isolated cells

总览

实验数据

产品说明书及文档

应用领域

总览

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

查看更多显示更少

实验数据

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.