Mutant Enrichment with 3′-Modified Oligonucleotides: A Practical PCR Method for Detecting Trace Mutant DNAs

作者: Seung-Tae Lee , Ji-Youn Kim , Min-Jung Kown , Sun Wook Kim , Jae Hoon Chung

DOI: 10.1016/J.JMOLDX.2011.07.003

关键词:

摘要: Many clinical situations necessitate highly sensitive and reliable molecular assays; however, the achievement of such assays remains a challenge due to inherent limitations testing methods. Here, we describe simple inexpensive enrichment technique that call mutant with 3′-modified oligonucleotides (MEMO). The method is based on use oligonucleotide primer blocks extension normal allele but enables mutated allele. performance was evaluated respect its ability detect common cancer mutations in EGFR, KRAS, BRAF, TP53, JAK2, NPM1 genes. We achieved sensitivities 10−2 10−6 using downstream Sanger sequencing, depending concentrations thermodynamics primers. MEMO may be applicable quantitative real-time PCR platform other assays. This practically various medical situations.

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