Uracil-DNA Glycosylase, heat-labile recombinant from marine bacterium BMTU 3346 
UNG with an increased heat intolerance to make it the solution of choice for the use in PCR carryover prevention.
Contents
The enzyme is supplied as 1 U/µl solution in storage buffer.
Application
U-DNA cleavage can be used to increase the efficiency of site-directed mutagenesis. For this purpose, uracil is incorporated in vivo into the target DNA. After primer annealing, and enzymatic filling-in, and ligation, the selection for the mutated strand is performed by enzymatic degradation of the original non-mutated uracil, which contains the wild-type strand, using UNG. A further application is carryover prevention for PCR. During PCR, dUTP can be incorporated into the PCR product. Before a subsequent amplification reaction, contaminating PCR products are degraded using uracil-DNA glycosylase. The enzyme is inactivated at the beginning of the amplification reaction by heating to 95°C. Simultaneous strand cleavage of the U-DNA is achieved during this heating step.
Background Information
Uracil-DNA glycosylase (UNG) hydrolyzes uracil-glycosidic bonds in single- or double-stranded DNA, excising uracil and creating alkali-sensitive abasic sites in the DNA. These abasic sites can be hydrolyzed by endonuclease, heat, or alkali treatment. dUTP-containing DNA (U-DNA) can be prepared by in vivo or in vitro methods. Depending on how the DNA is prepared, uracil-DNA glycosylase can be used to achieve general, site-specific, or strand-specific U-DNA cleavage. Ribouracil residues in RNA are a poor substrate for UNG.