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Cat. No.: HLUDG-500 (for 200U, 2 U/µl)

Cat. No.: HLUDG-1k (for 1000U, 20 U/µl)

For the standard version, please visit Uracil N-Glycosylase.

Description

Heat-Labile Uracil DNA Glycosylase (HL-UDG), also known as Antarctic ThermoLabile UDG, is a recombinant protein derived from psychrophilic marine bacteria and expressed and purified from E. coli. This enzyme efficiently hydrolyzes uracil residues in single-stranded or double-stranded DNA, creating abasic sites that easily break under high temperature or high pH. This ThermoLabile UDG has no activity on RNA and is primarily used to prevent PCR product contamination. Unlike the E. coli-derived uracil-DNA glycosylase, which is heat-stable and retains activity even after 10 minutes at 95ºC, potentially degrading PCR products containing dU bases, the ThermoLabile UDG from psychrophilic marine bacteria is completely inactivated after 5 minutes at 50ºC. To eliminate residual PCR product contamination, add ThermoLabile UDG to the PCR mix before amplification and incubate at 25ºC for 2 minutes. Since the enzyme is inactivated during the denaturation step at 94ºC in the PCR cycle, it does not affect the new PCR products containing dU.

Applications

• Removal of uracil bases from single-stranded or double-stranded DNA.

• Elimination of residual PCR contamination.

Activity Definition

One unit of activity is defined as the amount of enzyme required to catalyze the release of 60 pmol of uracil from uracil-containing double-stranded DNA per minute at 37ºC.

Reaction Buffer

ThermoLabile UDG is compatible with most PCR polymerase reaction buffers, but its activity is inhibited at high ionic concentrations (>100 mM).

Enzyme Storage Solution

20 mM Tris-HCl, 50 mM NaCl, 1 mM DTT, 50% Glycerol, 0.1% (w/v) Triton X-100, pH 7.5.

Storage

Store at -20ºC for up to 2 years.

Heat Inactivation

50ºC for 5 minutes.