T7 Endonuclease I

Unit Definition

In a 50 μL reaction system, at 37°C for 1 h, the amount of enzyme required to convert more than 90% of the pUC(AT)* of a 1-ug supercoiled cross-shaped structure into more than 90% of the linear structure is defined as an active unit.

Components

Application

• lDetection of mutant caused by gene mutation, SNP, TALEN or CRISPR/Cas9

• lRecognition of mismatched DNA

• lDecomposition of four-way junction DNA or Branched DNA

• lDetection or cutting of heterodimer DNA

• lShot-gun cloning by random cutting of linear DNA

1X T7 Endonuclease I Buffer

50 mM NaCl, 10 mM Tris-HCl, 10 mM MgCl2, 1 mM DTT (pH 7.9 at 25°C).

Preservation Reagent

50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5.

Storage

Stored at -20°C for 2 years. Avoid repeated freezing and thawing.

Notes

• lT7 endonuclease I is an enzyme with substrate structure selectivity. It acts on different DNA substrates with different activities. To cut a specific substrate, the amount of enzyme and reaction time must be carefully controlled.

• lWhen the reaction temperature exceeds 42°C, the non-specific nuclease activity will be increased.