ArciTect™ T7 Endonuclease I Kit
For estimation of CRISPR-Cas9 genome editing efficiency
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- Nuclease-Free Water
DNase- and RNase-free water for molecular biology applications
- Gel and PCR Clean-up Kit
For extraction of DNA from agarose gels, or clean-up of PCR products and restriction enzyme digestions
- Proteinase K Solution
For removing nucleases from DNA/RNA samples and terminating enzymatic reactions
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.
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Resources and Publications
Educational Materials (5)
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ArciTect™ T7 Endonuclease I Kit
The purchase of the ArciTect™ products conveys to the purchaser the limited, non-transferable right to use, in accordance with applicable laws and regulations, such purchased products and any related material solely for research purposes only, not for any commercial purposes, with no warranty from the owner or assignee of the Patents, and with no liability to the owner or assignee of the Patents, or a director, trustee, officer, employee, agent, faculty, official investigator or student thereof. Distribution of ArciTect™ products is covered under at least US 8,697,359, US 8,771,945, US 8,795,965, US 8,865,406, US 8,871,445, US 8,889,356, US 8,889,418, US 8,895,308, US 8,906,616 and foreign equivalents (“Patents”).
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