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제목 SNP genotyping through the melting analysis of unlabelled oligonucleotide applied on dilute PCR amplicon
등록일 2015-12-09 첨부파일
구분 SCI
학술지 JOURNAL OF BIOTECHNOLOGY
발표일 2011-07-20
저자 정상균, 유현주, 이영신, 김종열
Adaptation of DNA melting analysis for polymorphic single nucleotides (SNPs) genotyping using an unlabeled oligonucleotide probe for polymorphic DNAs under the presence of fluorescent DNA binding dye necessitates a reaction condition where the probe efficiently associates with a target strand that is PCR amplified. We present experimental evidence that application of an unlabeled probe to a dilute PCR amplicon provides a condition such that the fluorescent signals gained subsequently by probe melting are sufficient to discriminate allelic identities. This approach is best exploited by adapting the multiplexing PCR technique in order to cover multiple SNPs for given samples. 3′-end modification of the probe is unnecessary as the amplicon dilution step provides a way of inactivating the polymerase through divalent cation chelation. With the use of low-cost reagents and ordinary laboratory equipment, this method offers a rapid, simple and cost-efficient way of SNP genotyping.

*원문신청: kdc@kiom.re.kr

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