Contents > Molecular Cloning Enzymes > HK™-UNG Thermolabile Uracil N-Glycosylase

HK™-UNG Thermolabile Uracil N-Glycosylase

Supplier: EPICENTRE® Biotechnologies

HK™-UNG Thermolabile Uracil N-Glycosylase

Catalogue number

Pack size

HU5901K

1U/µl 1,000U

HU59100

1U/µl 100U

Note: This product is accompanied by a limited non-exclusive license for the purchaser to use the purchased product for Base Excision Sequence Scanning (BESS-T&G™ Scanning) in life science research under U.S. Patent No. 6,048,696 and similar patents pending in the U.S. and other countries. No other license for any other use or field of use is either granted or implied by the sale of this product. Also see Section 6 for information on BESS-T&G™ Base Reader Kits.

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Last modified: 2008-07-24 11:41:04

HK™-UNG is a Uracil N-Glycosylase that can be easily heat-inactivated. Uracil N-Glycosylase hydrolyses the N-glycosidic bond between the deoxyribose sugar and uracil in DNA containing deoxyuridine in place of thymidine. HK™-UNG is active on both single and double-stranded DNA that contains uracil, but has no activity on RNA or 2'-deoxyuridine-5'-monophosphate.

Unit Definition:

One unit catalyses the release of 1 nanomole of uracil from uracil-containing DNA in 1 hour at 37°C in 40mM Tris-HCl, pH 8.3, 1mM EDTA.
Note: A unit as defined by Epicentre® is 5- to 20-fold more active than a unit as defined by other suppliers. Therefore, Dilution Buffer is provided for applications requiring lower concentrations of enzyme.

Dilution and Storage Buffers:

50mM Tris-HCl, pH 7.5, 100mM NaCl, 0.1mM EDTA, 1mM DTT, and 0.1% Triton® X-100 in 50% glycerol.

Quality Control:

HK™-UNG is free of detectable exo- and endonuclease, and RNase activities.

Last modified: 2008-05-14 16:46:46

Benefits

HK™-UNG is fully active at 50°C, but is inactivated by a 10-minute incubation at 65°C
High activity - requires only 0.1 unit in a standard reaction

Figure 1. Brief heat treatment of HK™-UNG eliminates its ability to digest dU-containing DNA. One unit of each uracil N-glycosylase was incubated for 10 minutes at the indicated temperature in 20µl of a Tris-acetate buffer (pH 8.3). Following heat treatment, ≈300 ng of a dU-containing, 450-bp DNA fragment were added and the samples were incubated at 37°C for 30 minutes. Five µl of a stop buffer were added and the samples were separated in a 1.4% agarose gel. The gel was stained with ethidium bromide and the DNA was visualized under UV illumination. The presence of the 450-bp band in Lanes 5 and 7 demonstrates that HK-UNG is inactivated at 70°C and 95°C.