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T7 Endonuclease I (T7E1) recognizes and cleaves non-perfectly matched DNA, cruciform DNA structures, Holliday structures or junctions, hetero duplex DNA and more slowly, nicked double-stranded DNA. The cleavage site is at the first, second or third phosphodiester bond that is 5´ to the mismatch. The protein is the product of T7 gene 3. GenCrispr T7 Endonuclease I is a fusion protein produced from E.coli.
Synonyms
T7E1
Note
T7 Endonuclease I is a structure-selective enzyme. It acts on a variety of DNA substrates with different specific activities. It is important to control the amount of enzyme and the reaction time used for cleavage of a particular substrate. Temperatures above 42°C cause an increase in nonspecific nuclease activity and should be avoided.
Applications
Resolve four-way junction or branched DNA.
Detect or cleave hetero duplex and nicked DNA.
Randomly cleave linear DNA for shot-gun cloning.
Detect gene mutagenesis and SNPs, for cleavage efficiency assays induced by TALEN,CRISPR/CAS9 or other gene editing tools.
Properties
Storage & Stability
GenCrispr T7 Endonuclease I is supplied in 1X storage buffer (200 mM NaCl,20 mM Tris-HCl(pH 7.5), 0.1 mM EDTA, 1 mM dithiothreitol, 0.15% Triton X-100 and 50% glycerol). The recommended storage temperature is -20°C.