Takara TaKaRa PCR Carryover Prevention Kit

Carryover of amplification products from previous PCR reactions may lead to false-positive results. This is a challenge when using endpoint PCR for food and environmental sample testing. The TaKaRa PCR Carryover Prevention Kit is designed to preclude false-positive results caused by carryover contamination. This kit includes dUTP instead of dTTP for PCR, and includes UNG (uracil-N-glycosylase), an enzyme that degrades uracil-containing DNA.

Carryover of amplification products from previous PCR reactions may lead to false-positive results. This is a challenge when using endpoint PCR for food and environmental sample testing. The TaKaRa PCR Carryover Prevention Kit is designed to preclude false-positive results caused by carryover contamination. This kit includes dUTP instead of dTTP for PCR, and includes UNG (uracil-N-glycosylase), an enzyme that degrades uracil-containing DNA. Treatment with UNG prior to PCR degrades uracil-containing PCR contaminants from prior PCR reactions but leaves thymine-containing templates intact, allowing selective removal of carryover PCR products. The kit contains the enzyme UNG, dNTP mix containing dUTP, and MgCl2 solution.

Degradation by UNG takes place in the following manner. In a pre-PCR reaction at 25˚C for 10 min, UNG hydrolyzes N-glycosylic bonds between the deoxyribose sugars and the uracil bases in uracil-containing DNA, leaving apyrimidinic sites in the DNA. Next, a heat treatment at 95˚C for 2 min deactivates UNG and simultaneously causes cleavage and degradation of the contaminant DNA fragments by hydrolysis of the phosphate backbone at the abasic sites. Use this kit in combination with Takara Taq hot-start version (Cat. # R007A/B) or other Pol I-type PCR enzymes.

UNG hydrolyzes uracil-containing single- and double-stranded DNA but shows no activity toward RNA.

More Less