Thermo Scientific™ Uracil-DNA Glycosylase (1 U/μL)

Description

Uracil-DNA Glycosylase (UDG, UNG) catalyzes the hydrolysis of the N-glycosylic bond between uracil and sugar, leaving an apyrimidinic site in uracil-containing single or double-stranded DNA. The enzyme shows no activity on RNA.

• Active in Fermentas buffers for restriction enzymes and thermophilic polymerases
• Source: E. coli K12 cells
• Molecular Weight: 25.6 kDa monomer

• The absence of endo-, exodeoxyribonucleases, phosphatases and ribonucleases confirmed by appropriate quality tests

Source:

• E.coli K12 cells

Molecular Weight:

• 25.6kDa monomer

Definition of Activity Unit:

• One unit of the enzyme catalyzes the release 1 nanomole of uracil from uracil-containing DNA template in 60 min. at 37°C

• 30mM Tris-HCl (pH 7.5), 150mM NaCl, 1mM EDTA, 1mM DTT, 0.05% (v/v) Tween 20 and 50% (v/v) glycerol

10X Reaction Buffer:

• 200mM Tris-HCl (pH 8.2 at 25°C), 10mM EDTA, 100mM NaCl

• Inhibitors: Ugi protein from the Bacillus subtilis phage PBS2, protein p56 from the Bacillus subtilis phage phi29 (7).
• Inactivated by heating at 95°C for 10 min. Enzyme activity is partially restored at temperatures lower than 55°C. Therefore put PCR products on ice after PCR and load directly on a gel

Recommended for:

Control of carry-over contamination in PCR (2); Glycosylase mediated single nucleotide polymorphism detection (GMPD) (3); Site-directed mutagenesis (4); As a probe for protein-DNA interaction studies (5); SNP genotyping; Cloning of PCR products (6); Generation of single strand overhangs of PCR products and cDNA

Note:

The abasic sites formed in DNA by Uracil-DNA Glycosylase may be subsequently cleaved by heat, alkali-treatment or endonucleases that cleave specifically at abasic sites.UDG (UNG) is active in the presence or absence of divalent cations.