T7E1 CRISPR Validation Kit by Merck

Manufacturer Merck  |  Available in Asia, Australia Oceania
The T7 Endonuclease Detection Assay is a well-known method for detecting genome editing events from CRISPR, Zinc-finger nuclease, and TALEN gene targeting. Originally identified from Escherichia coli bacteriophage, the T7 endonuclease can cleave mismatched heteroduplex DNA, Holliday junctions, branched DNA, and cruciform DNA.


T7E1 CRISPR Validation Kit by Merck product image
T7E1 CRISPR Validation Kit
Request Pricing

Merck

Receive your quote directly from the manufacturer.



0 Scientists have reviewed this product


Write the First Review

No Reviews

Following a gene editing experiment, genomic DNA surrounding the target locus is amplified by PCR, and the PCR amplicons are denatured and reannealed through heating and slow cooling. If NHEJ events have occurred, then, after reannealing, several products are possible.

Homoduplexes can form where a WT strand is reannealead to a WT strand or an indel-carrying strand is reannealed to an indel-carrying strand. Heteroduplexes form when a WT strand is reannealed to an indel-carrying strand causing a mismatch. Heteroduplex products with mismatches are cleaved by the T7 endonuclease. Separating the DNA products after treatment with T7 endonuclease by gel electrophoresis will result in a banding pattern indicative of the amount of heteroduplexes in the sample. The amount of cleaved heteroduplexes is directly related to the amount of indel activity.

Application

Functional Genomics; Target Validation; Genome Editing

Features and Benefits

  • Technically simple method based on well-known techniques
  • Easily interpretable results
  • Fast analysis turnaround
  • Cost-effective