RNase H and its effects on PCR

摘要

RNase H was originally described as an enzyme that specifically degrades RNA in an RNA:DNA hybrid and is inactive on ss- or dsRNAs and DNAs alone (2). RNase H treatment is recommended during cDNA preparation because the RNase H activity, originally present in reverse transcriptases, has been removed in some cloned enzymes. These modified enzymes yield more cDNA, so they are now more routinely used in the preparation of cDNA. It is believed that low copy number transcripts may be more readily detected after RNase H treatment, but systematic studies demonstrating the effects of RNase H have not been reported. We show that the template of one gene of a multigene family becomes more readily available for PCR amplification only after RNase H treatment, while two other genes are not affected by RNase H treatment.