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RNase H2-dependent PCR (rhPCR) is a powerful method for increasing PCR specificity and eliminating primer dimers by using blocked primers and a thermostable RNase H2 from Pyrococcus abyssi. Primers will only support extension and replication after the blocked portion is cleaved. Cleavage by the RNase H2 enzyme occurs only when primers are bound to their complementary target sequence, thus providing increased specificity. Also, the thermostability of P. abyssi RNase H2 provides a “hot start” capability to the reaction. In this presentation, Dr Joseph Dobosy (senior research scientist in the molecular genetics research division of IDT) gives a detailed explanation of the rhPCR mechanism, offers tips on how to design assays using this powerful technology, and provides examples of applications that benefit from rhPCR.