Figure 1.

Coupled reaction scheme for PCR using blocked primers activated by cleavage with RNase H2 (rhPCR). PCR primers are designed to be incapable of extension by DNA polymerase and contain a single ribonucleotide residue near the 3'-end. Hybridization of primer to template forms a substrate for RNase H2, which will cleave the primer 5'-to the RNA base leaving a DNA oligonucleotide with a 3'-OH capable of priming DNA synthesis. The assay can be performed using either 2- or 3-step PCR with anneal/extend times as short as 30 seconds.

Dobosy et al. BMC Biotechnology 2011 11:80   doi:10.1186/1472-6750-11-80
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