EnGen Mutation Detection Kit, New England Biolabs

Supplier: New England Biolabs
E3321S E3321S
CA103049-174EA 342 CAD
103049-174 CA103049-174
EnGen Mutation Detection Kit, New England Biolabs
Electrophoresis Systems Mutation Detection Systems
The EnGen Mutation Detection Kit provides reagents for detection of on-target genome editing events.

In the first step, targeted regions from cells whose genomes were targeted (i.e. CRISPR/Cas9, TALENs, Zinc-finger Nucleases) are amplified using Q5 Hot Start High-Fidelity 2X Master Mix. Upon denaturation and re-annealing, heteroduplexes are formed when mutations from insertions and deletions (indels) are present in the amplicon pool. In the second step, annealed PCR products are digested with EnGen T7 Endonuclease I, a structure-specific enzyme that will recognize mismatches larger than 1 base. Both strands of the DNA are cut when a mismatch is present, which results in the formation of smaller fragments. Analysis of the resulting fragments provides an estimate of the efficiency of the genome editing experiments.
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