Fluorescence-based DHPLC for allelic quantification by single-nucleotide primer extension

Kenjiro Kosaki, Hiroshi Yoshihashi, Yuko Ohashi, Rika Kosaki, Taichi Suzuki, Nobutake Matsuo

研究成果: Article査読

7 被引用数 (Scopus)

抄録

We have investigated the possibility of determining quantitatively the alleles of binary DNA polymorphisms by single-nucleotide primer extension (SNuPE) and fluorescence-based DHPLC. Using a polymorphism of interest to our group, ROX-labeled dideoxy CTP (ROX-ddCTP) was incorporated at the 3′ end of the primer annealed to the template adjacent to the polymorphic site. The primer extension product was then resolved from the unincorporated dye terminator by ion-pair reversed-phase liquid chromatography. The signal intensity of incorporated ROX-ddCTP correlated well over one order of magnitude with the relative amount of the C-allele present in the genomic DNA template. We conclude that SNuPE, when combined with fluorescence-based DHPLC, can accurately determine the relative molar proportion of one allele in total DNA.

本文言語English
ページ(範囲)111-119
ページ数9
ジャーナルJournal of Biochemical and Biophysical Methods
47
1-2
DOI
出版ステータスPublished - 2001 1月 30

ASJC Scopus subject areas

  • 生物理学
  • 生化学

フィンガープリント

「Fluorescence-based DHPLC for allelic quantification by single-nucleotide primer extension」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

引用スタイル