RNase H-dependent PCR (rhPCR) (CAT#: STEM-MB-0217-WXH)

Introduction

RNase H-dependent PCR (rhPCR) is a nucleic acid amplification method that provides increased target specificity over traditional PCR. Compared to traditional PCR, rhPCR requires an additional enzyme, RNase H2, and uses blocked primers in place of conventional PCR primers.<br />In rhPCR, the primers are designed with a removable amplification block on the 3’ end. Amplification of the blocked primer is dependent on the cleavage activity of a hyperthermophilic archaeal Type II RNase H enzyme during hybridization to the complementary target sequence. This RNase H enzyme possesses several useful characteristics that enhance the PCR. First, it has very little enzymatic activity at low temperature, enabling a “hot start PCR” without modifications to the DNA polymerase. Second, the cleavage efficiency of the enzyme is reduced in the presence of mismatches near the RNA residue. This allows for reduced primer dimer formation, detection of alternative splicing variants, ability to perform multiplex PCR with higher numbers of PCR primers, and the ability to detect single-nucleotide polymorphisms.<br />The primer-dimer reducing abilities of rhAmp technology make it ideal for demanding genomics applications like multiplexed amplicon sequencing and SNP genotyping.