LightCycler® Uracil-DNA Glycosylase hydrolyzes uracil-glycosidic bonds at U-DNA sites in single- and double-stranded DNA, excising uracil and creating alkali- and heat-sensitive abasic sites in the DNA.
- The enzyme is more active on single-stranded DNA than on double- stranded DNA.
- The enzyme is inactive on RNA and native, uracil-free DNA.
The enzyme is supplied in storage buffer at 2 U/µl.
LightCycler® Uracil-DNA Glycosylase (LightCycler® UNG) is ideally suited for preventing carryover contamination between PCRs. This technique involves incorporating deoxyuridine triphosphate (dUTP) into all amplification reactions, and pretreating all successive LightCycler® FastStart Enzyme-based PCR mixtures with LightCycler® UNG. Prior to the thermal-cycling procedure, LightCycler® UNG is added to the LightCycler® amplification reaction mixture, then incubated for 10 minutes at +40°C. During the initial activation of FastStart DNA Polymerase (+95°C for 10 minutes), the LightCycler® UNG becomes inactive.
Note: LightCycler® UNG is designed only for use with the FastStart Enzyme or FastStart Enzyme-based LightCycler® Kits.
Function test: LightCycler® UNG is function-tested for carryover prevention using the LightCycler® Carousel-Based System.