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Cat. No.: DFNTH-500 (for 500U)

Cat. No.: DFNTH-2500 (for 2500U)

Description

E. coli Endonuclease III (Nth) exhibits both N-glycosylase and AP-lyase activities. The N-glycosylase activity releases damaged pyrimidine bases from double-stranded DNA, creating an apyrimidinic (AP) site. The AP-lyase activity cleaves the 3’ end of the AP site, generating a 5’ phosphate and a 3’ phospho-α,β-unsaturated aldehyde.

The damaged bases recognized and excised by Endonuclease III include:

• Urea
• 5,6-dihydroxythymine
• Thymine glycol
• 5-hydroxy-5-methylhydantoin
• Uracil glycol
• 6-hydroxy-5,6-dihydroxythymine
• Methyleneglycolhydantoin
• Production

Our DNA-Free E. coli Endonuclease III (Nth), on the other hand, is devoid of genomic and plasmid DNA from the production strain. This makes DNA-Free E. coli Endonuclease III (Nth) particularly suitable for amplifying, sequencing, and cloning 16S and 23S rDNA genes, as well as for standard applications like PCR and primer extension.

Activity Definition

One unit of activity is defined as the amount of enzyme required to cleave 1 pmole of a 34 bp oligonucleotide duplex containing a single AP site in 1 hour at 37°C in a 1× Endonuclease III reaction buffer.

Activity Assay Conditions

1× Endonuclease III reaction buffer: 20 mM Tris-HCl (pH 8.0), 1 mM EDTA, 1 mM DTT, incubated at 37°C.

Applications

• Single-cell gel electrophoresis (Comet assay)
• DNA damage repair

Storage

Store at -20°C for up to 2 years. Avoid repeated freeze-thaw cycles.