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U1900-05 Uracil DNA Glycosylase (UNG) CAS:

Specifications
References
Grade
Molecular Biology Grade
Molecular Weight
25.6
EU Commodity Code
38220090
Shipping Temp
Blue Ice
Storage Temp
-20°C
Supplied with 10X Reaction buffer

Catalyzes the hydrolysis of N-glycosylic bond between the uracil and sugar, leaving an apyrimidinic site in uracil-containing single or double-stranded DNA. It shows no activity on RNA (1).

Unit Definition
1unit of Uracil DNA Glycosylase catalyses the release of 1nanomole of uracil from uacil containing DNA template in 60 min at 37°.
Storage Buffer
Supplied as a liquid in 30mM Tris-HCl, pH 7.5, 150mM sodium chloride, 1mM EDTA, 1mM DTT, 0.05% Tween 20, 50% glycerol.
10X Reaction Buffer (U1900-05A)
Supplied as a liquid in 200mM Tris-HCl, pH 8.2, 10mM EDTA, 100mM sodium chloride.
Applications
Suitable for GMPD detection, Site-directed mutagenesis (2), as a Probe for protein-DNA interaction (3), Rapid, efficient cloning of PCR products (4) anddH2Oelimination of carry over contamination in PCR.
Note
The abasic sites formed in DNA by uracil-DNA glycosylase may be cleaved by heat under alkaline conditions. Any of the five USBio buffers can be used instead of 10X uracil-DNA glycosylase reaction buffer. Uracil-DNA glycosylase should be inactivated by heating at 95°C for 10 minutes. Enzyme activity is partially restored at temperatures lower than 55°C. Put PCR products on ice after PCR and load directly on a gel.
Inhibitors
Ugi protein from the Bacillus subtilis phage PBS2, protein p56 from the Bacillus subtilis phage phi29 (7). UGI is active in the presence or absence of divalent cations.
Endodeoxyribonuclease Assay
No detectable conversion of covalently closed circular DNA to nicked DNA was observed after incubation of 10u of Uracil DNA Glycosylase with 1ug of pUC19 DNA in 50ul of reaction buffer (33mM Tris- acetate (pH7.9 at 37ºC), 10mM Mg-acetate, 66mM K-actetate) for 4 hours at 37°C.
Labeled Oligonucleotide Assay
No detectable degradation of single- stranded and double-stranded labeled oligonucleotide was observed after incubation with 2u of Uracil DNA Glycosylase, for 4 hours at 37°C.
Ribonuclease Assay
≤0.5% of the total radioactivity was released into trichloroacetic acid-soluble fraction after incubation of 10 units of Uracil DNA Glycosylase with 1ug of [3H]-RNA in 50ul of reaction buffer (33mM Tris- acetate (pH7.9 at 37ºC), 10mM Mg-acetate, 66mM K-actetate) for 4 hour at 37°C.
Storage and Stability
Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Source
E. coli strain K12
Concentration
~1u/ul
Important Note
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
References
1. Lindahl, T. et al., (1977) Biol. Chem. 252(10):3286–3294. 2. Kunkel, I.A., (1985) Proc. Natl. Acad. Sci. USA 82:488–492. 3. Devchand, P.R., et al., (1993) Nucleic Acids Res. 21:3437–3443. 4. Booth, P.M., et al., (1994) Gene 146:303–308.
USBio References
No references available
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