Uracil-DNA Glycosylase
Detailní popis
Thermo Scientific 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 (see Figure 1 in Supporting Data). The enzyme shows no activity on RNA (see Reference 1).
Highlights
- Active in Thermo Scientific buffers for restriction enzymes and thermophilic polymerases
Applications
- Control of carry-over contamination in PCR (see Reference 2)
- Glycosylase mediated single nucleotide polymorphism detection (GMPD) (see Reference 3)
- Site-directed mutagenesis (see Reference 4)
- As a probe for protein-DNA interaction studies (see Reference 5)
- SNP genotyping
- Cloning of PCR products (see Reference 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.
Use of this enzyme in certain applications may be covered by patents and may require a license.
10X Reaction Buffer | 200 mM Tris-HCl (pH 8.2 at 25°C), 10 mM EDTA, 100 mM NaCl |
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Concentration | 1 U/µL |
Definition of Activity Unit | One unit of the enzyme catalyzes the release 1 nm of uracil from uracil-containing DNA template in 60 min at 37°C. |
Hazardous | No |
Hazardous: | No |
Inactivation | 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. |
Inhibition | Inhibitors: Ugi protein from the Bacillus subtilis phage PBS2, protein p56 from the Bacillus subtilis phage phi29 (see Reference 7). |
Molecular Weight | 25.6 kDa monomer |
Quality Control | The absence of endo-, exodeoxyribonucleases, phosphatases, and ribonucleases confirmed by appropriate quality tests. |
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Shipping Information | |
Source | E. coli K12 cells |
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Storage Buffer | The enzyme is supplied in 30 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EDTA, 1 mM DTT, 0.05% (v/v) Tween 20, and 50% (v/v) glycerol. |
Storage Condition | -20 C |
Storage Condition: | -20 C |
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