Recombinant E. coli ung Protein

Cat.No.: CMC13
Product Overview: Recombinant ung protein was expressed in an E. coli strain that carries the UDG gene from E. coli.
Description: UDG and Ndk form a complex (Goswami, 2006). [More information is available at EcoGene: EG11058]. Uracil can be generated in DNA as a result of misincorporation by DNA polymerase, spontaneous deamination of cytosine, and deamination of cytosine induced by bisulfite or nitrous acid.
Source: E. coli
Species: E. coli
Form: Reagents Supplied
The following reagents are supplied with this product:
10X UDG Reaction Buffer
Unit Definition: One unit is defined as the amount of enzyme that catalyzes the release of 60 pmol of uracil per minute from double-stranded, uracil-containing DNA. Activity is measured by release of [3H]-uracil in a 50 µl reaction containing 0.2 µg DNA (10^4-10^5 cpm/µg) in 30 minutes at 37 centigrade.
Applications: Treatment of 0.1 µg of uracil-containing DNA with 1 unit of UDG for 10 minutes at 37 centigrade renders the DNA incapable of being copied by DNA polymerase. The enzyme can be 95% heat killed by incubation at 95 centigrade for 10 minutes. Since UDG remains partially active following heat treatment at 95 centigrade, it is recommended that uracil glycosylase inhibitor be added to prevent degradation of product DNA. Alternatively, reaction products can be immediately extracted with phenol/chloroform.
Usage: Reaction Conditions
1X UDG Reaction Buffer
Incubate at 37 centigrade

1X UDG Reaction Buffer
20 mM Tris-HCl
1 mM DTT
1 mM EDTA
(pH 8 @ 25 centigrade)

Heat Inactivation
No

Unit Assay Conditions
1X UDG Reaction Buffer, 1 unit of uracil DNA Glycosylase, 0.2 µg 3H-uracil DNA (10^4-10^5 cpm/μg) for 30 minutes at 37 centigrade in a total reaction volume of 50 µl.
Notes: UDG is active over a broad pH range with an optimum at pH 8.0, does not require divalent cation, and is inhibited by high ionic strength (> 200 mM).
Storage: At -20 centigrade.
Concentration: 5,000 units/mL
Storage Buffer: 10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
0.1 mg/ml BSA
50% Glycerol
pH 7.4 @ 25°C
Gene Name: ung
Official Symbol: ung uracil-DNA-glycosylase [ Escherichia coli str. K-12 substr. MG1655 ]
Synonyms: ung; uracil-DNA-glycosylase; ECK2578; JW2564; uracil-DNA-glycosylase; EC 3.2.2.-
Gene ID: 947067
Protein Refseq: NP_417075
UniProt ID: P12295

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