ArciTect™ T7 Endonuclease I Kit

For estimation of CRISPR-Cas9 genome editing efficiency
Catalog #
76021_C
For estimation of CRISPR-Cas9 genome editing efficiency
From: 79 USD
Required Products
  1. Nuclease-Free Water|79001
    Nuclease-Free Water

    DNase- and RNase-free water for molecular biology applications

  2. Genomic DNA Purification Kit
    Genomic DNA Purification Kit

    For purification of genomic DNA from cells or tissue

  3. Proteinase K Solution
    Proteinase K Solution

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

  4. 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

Overview

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.
Components
ArciTect™ T7 Endonuclease I Kit, 25 Reactions (Catalog #76021)
• ArciTect™ T7 Endonuclease I, 25 Reactions
• ArciTect™ T7 Endonuclease I Buffer (10X), 1 mL
 
ArciTect™ T7 Endonuclease I Kit, 125 Reactions (Catalog #76022)
• ArciTect™ T7 Endonuclease I, 125 Reactions
• ArciTect™ T7 Endonuclease I Buffer (10X), 1 mL
Cell Type
Pluripotent Stem Cells
Species
Human
Brand
ArciTect
Area of Interest
Stem Cell Biology

Scientific Resources

Product Documentation

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

Educational Materials (5)

Brochure
Products for Human Pluripotent Stem Cells
Technical Bulletin
Genome Editing with Direct Cas9 RNP Delivery Design Considerations
Technical Bulletin
Evaluation of Genome Editing
Technical Bulletin
Genome Editing of Human Primary T Cells Using CRISPR-Cas9
Technical Bulletin
Genome Editing of Human Pluripotent Stem Cells Using the ArciTect™ CRISPR-Cas9 System

Product 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.

Data and Publications

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.

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