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Detection of rare point mutation via allele-specific amplification in emulsion PCR
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  • Detection of rare point mutation via allele-specific amplification in emulsion PCR
  • Detection of rare point mutation via allele-specific amplification in emulsion PCR
저자명
Cheng. Changming,Zhou. Yin,Yang. Chao,Chen. Juan,Wang. Jie,Zhang. Jie,Zhao. Guoping
간행물명
BMB reports
권/호정보
2013년|46권 5호|pp.270-275 (6 pages)
발행정보
생화학분자생물학회
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정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

It is essential to analyze rare mutations in many fields of biomedical research. However, the detection of rare mutations is usually failed due to the interference of predominant wild-type DNA surrounded. Herein we describe a sensitive and facile method of detecting rare point mutation on the basis of allele-specific amplification in emulsion PCR. The identification and selective amplification of rare mutation are accomplished in one-pot reaction. The allele-specific primers coupled on magnetic beads allow the exclusive amplification and enrichment of the mutant amplicons. The productive beads bearing mutant amplicons are subsequently stained with the fluorescent dyes. Thus, the rare point mutations with a percentage as low as 0.1%, can be detected by fluorescent analysis. The relative percentages of mutation among different samples can be roughly accessed by counting the fraction of fluorescent positive beads through flow cytometry.