Science China Chemistry

, Volume 57, Issue 7, pp 961–965 | Cite as

RNA-primed allele-specific PCR

  • LingHui Zhang
  • Zhuo Tang


RNA/DNA primer pairs and two polymerases were used to efficiently amplify DNA sequences using the conventional polymerase chain reaction (PCR). The reaction required the use of both DNA polymerase and reverse transcriptase during each thermal cycle and formed a double-stranded DNA in which one terminus was an RNA/DNA hybrid. Because there is a higher sensitivity of the DNA polymerase to the mismatch at the 3′-end in the RNA/DNA hybrid duplex than in the DNA/DNA duplex, the RNA-primed PCR reveals much better specificity in the allele-specific PCR to detect single-nucleotide mutation.


RNA primer Vent(exo-) DNA polymerase TaqM1 DNA polymerase allele-specific PCR single-nucleotide mutation 


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Supplementary material

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Copyright information

© Science China Press and Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  1. 1.Natural Products Research Center, Chengdu Institute of BiologyChinese Academy of SciencesChengduChina
  2. 2.University of Chinese Academy of SciencesBeijingChina

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