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architect™T7核酸内切酶I检测试剂盒

用于估计CRISPR-Cas9基因组编辑效率
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产品号 #76021_C

用于估计CRISPR-Cas9基因组编辑效率

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Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.
  1. ArciTect™ High-Fidelity DNA Polymerase Kit
    ArciTect™ High-Fidelity DNA Polymerase Kit

    Polymerase, buffers, and dNTPs for high-fidelity PCR amplification

  2. Nuclease-Free Water
    Nuclease-Free Water

    DNase- and RNase-free water for molecular biology applications

  3. Proteinase K Solution
    Proteinase K Solution

    For removing nucleases from DNA/RNA samples and terminating enzymatic reactions

  4. Genomic DNA Purification Kit
    Genomic DNA Purification Kit

    For purification of genomic DNA from cells or tissue

  5. Gel and PCR Clean-up Kit
    Gel and PCR Clean-up Kit

    For extraction of DNA from agarose gels, or clean-up of PCR products and restriction enzyme digestions

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实验数据
产品说明书及文档
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概述

architect™T7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。architect™T7核酸内切酶I试剂盒由architect™T7核酸内切酶I和architect™T7核酸内切酶I缓冲液（10X）组成，它们已经过测试和验证，可用于architect™CRISPR-Cas9基因组编辑系统。architect™T7内切酶I识别和切割错配DNA、十字形DNA结构、Holliday结构或连接、异双工DNA，以及效率较低的缺口双链DNA。由于切割效率与在特定DNA靶标上创建的INDELs数量成正比，因此使用architect™T7内切酶I Kit以快速和经济的方式估计基因编辑效率

Cell Type
Pluripotent Stem Cells
 
Species
Human
 
Application
Genome Editing
 
Area of Interest
Stem Cell Biology
 

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Data Figures

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.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Information Sheet
Product Name
ArciTect™ T7 Endonuclease I Kit
Catalog #
76021, 76022
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
ArciTect™ T7 Endonuclease I Kit
Catalog #
76021, 76022
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
ArciTect™ T7 Endonuclease I Kit
Catalog #
76021, 76022
Lot #
All
Language
English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Research Area
Workflow Stages

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Species Human
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