〈論文・報告〉ホスホロチオエート核酸をプライマーとする一塩基変異識別定量PCR法

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  • 〈PAPERS and REPORTS〉Quantitative PCR Discriminating Single Base Mutation Using Phosphorothioate Primers

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In the present study, quantitative analysis of single nucleotide mutated genes by allele specific real time PCR using phosphorothioate primers and DNA polymerases is described. DNA polymerases involve taq DNA polymerases （TaKaRa Taq PolymeraseTM and HiDi PolymeraseTM） which do not have 3’-exonuclease activity and HiFi taq DNA polymerase （TaKaRa Ex Taq PolymeraseTM） which has 3’-exonuclease activity, namely proofreading activity. Consequently, accuracy of discrimination of single base mutation was not so high in the PCR using TaKaRa Taq PolymeraseTM and TaKaRa Ex Taq PolymeraseTM. On the other hand, HiDi PolymeraseTM showed high accuracy of discrimination of single base mutation between KRAS wt and KRAS G12D model DNA templates.

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