ArciTect™ T7 Endonuclease I Kit

For estimation of CRISPR-Cas9 genome editing efficiency

ArciTect™ T7 Endonuclease I Kit

For estimation of CRISPR-Cas9 genome editing efficiency

From: 86 USD
Catalog #
76021_C
For estimation of CRISPR-Cas9 genome editing efficiency

What's Included

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
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.

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.
Cell Type
Pluripotent Stem Cells
Species
Human
Brand
ArciTect
Area of Interest
Stem Cell Biology

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
Catalog #
76021, 76022
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
76021, 76022
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
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.

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