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Cat. No.: T7E1-250 (for 250U)

Cat. No.: T7E1-1250 (for 1250U)

Cat. No.: T7E1-5k (for 5000U)

Description

T7 Endonuclease I (T7 Endo I, T7EI) is a relatively unique DNA endonuclease that can recognize and cleave non-perfectly matched DNA, cruciform DNA structures, Holliday structures or junctions, and heteroduplex DNA. T7EI cleaves the sites located at the first, second, or third phosphodiester bond at the 5' end of mismatched bases. Additionally, T7EI can also cleave nicked double-stranded DNA at a slow rate. This product is commonly used for identifying gene mutations induced by CRISPR/Cas9 and other methods.

It is important to note that T7EI can recognize mismatches in DNA caused by insertions, deletions, or mutations of length greater than or equal to 2 base pairs (bp) but cannot recognize 1-bp insertions, deletions, or mutations.

Application

T7 Endonuclease I (T7 Endo I, T7EI) is commonly used for detecting mutations induced by CRISPR/Cas9, TALENs, or other gene editing tools. It is also utilized for the detection of gene point mutations and single nucleotide polymorphisms (SNPs). Furthermore, T7EI can be employed to cleave four-way junctions or branched DNA, as well as to detect or cleave heteroduplex and nicked DNA. Additionally, it is used for random cleavage of linear DNA, which involves partially paired double-stranded DNA formed within or between linear DNA molecules, for applications such as shotgun cloning and sequencing.

Source

Recombinant protein of T7 bacteriophage gene 3 expressed in Escherichia coli.

Activity Definition

One unit is defined as the amount of enzyme required to convert > 90% of 1 μg of supercoiled cruciform pUC(AT) DNA to > 90% linear form in a total reaction volume of 50 μl within 1 hour at 37°C.

10X Reaction Buffer

500 mM NaCl, 100 mM Tris-HCl, 100 mM MgCl2, 10 mM DTT, pH 7.9.

Purity

Purity is greater than 95%. The enzyme does not contain any DNA endonuclease or exonuclease other than T7 Endonuclease I, RNAse, or phosphatases.

Inactivation or Inhibition

T7EI can be inactivated by heating at 85°C for 15 minutes or by adding EDTA to a final concentration of 20 mM.