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Cat. No.: TENN4-500 (for 500U)

Cat. No.: TENN4-2500 (for 2500U)

The lyophilized Thermostable Endonuclease IV is also available, which can be transported at room temperature.

Description

Thermostable Endonuclease IV, derived from thermophilic bacteria, is a nucleic acid endonuclease with specificity for apurinic/apyrimidinic (AP) sites. The enzyme cleaves the first phosphodiester bond at the 5´ end of the AP site, generating a 3´ hydroxyl and a 5´ deoxyribose-phosphate terminus. In addition to natural AP sites, various synthetically engineered spacer molecules can be effectively cleaved by endonuclease IV, including THF, Spacer C3, Spacer C12, Spacer 9, Spacer 18, and others. The enzyme also exhibits 3´-phosphodiesterase activity, releasing phosphoglycoaldehyde, α,β-unsaturated aldehydes, 3´-phosphate, and other 3' blocking groups from the 3´ end of DNA. Moreover, it acts on several oxidative damages on DNA molecules, hydrolyzing the first phosphodiester bond at the 5' end of oxidatively damaged bases. Additionally, the enzyme possesses 3´-5´ exonuclease activity. Its activity is influenced by metal ions, DTT, EDTA, and it preferentially acts on the 3´ recessed ends of double-stranded DNA.

Activity Definition

One unit is defined as the amount of enzyme required to cleave 1 pmol of a 34 bp fluorescently labeled oligonucleotide double-stranded DNA containing a single AP (apurinic/apyrimidinic) site in 1 hour at 65°C.

The preparation method for the AP site is as follows: Under 37°C conditions, treat a 34 bp oligonucleotide double-stranded DNA containing one uracil base with 1 unit of uracil-DNA glycosylase (UDG) for 2 minutes.

Activity Assay

1× AM Buffer E: 10 mM KCl, 20 mM Tris-HCl, 10 mM (NH4)2SO4, 2 mM MgSO4, 0.1% Triton X-100 (pH 8.8 @ 25°C), incubate at 65°C.

Concentration

10 U/μl

Features

• Exhibits excellent thermal stability, active in the temperature range of 65-75°C.

• Active in various reaction systems.

Applications

• Improves the performance of high-fidelity DNA polymerase PCR amplification when used in conjunction with a Thermostable UDG.

• Single-cell gel electrophoresis (Comet assay).

• Cleavage of various Spacers in synthesized oligonucleotides, such as Spacer C3, Spacer C18, Spacer 9, etc.

• Repair of damaged DNA.

Storage

Store at -20°C.

Only for research and not intended for treatment of humans or animals