PrimRglo: A multiplexable quantitative real-time polymerase chain reaction system for nucleic acid detection

Richard Lai, Fang Liang, Darnley Pearson, Graeme Barnett, David Whiley, Theo Sloots, Ross T. Barnard, Simon R. Corrie

Research output: Contribution to journalArticleResearchpeer-review

4 Citations (Scopus)

Abstract

We report the development of a new real-time polymerase chain reaction (PCR) detection system that uses oligonucleotide “tagged” PCR primers, a fluorophore-labeled “universal” detection oligonucleotides, and a complementary quenching oligonucleotide. The fluorescence signal decreases as PCR product accumulates due to the increase in detection/quencher hybrid formation as the tagged primer is consumed. We use plasmids containing the influenza A matrix gene and the porA and ctrA genes of Neisseria meningitidis as targets for developing the system. Cycle threshold (Ct) values were generated, and the sensitivity of the new system (dubbed “PrimRglo”) compared favorably with the commonly used SYBR green and Taqman detection systems and, unlike the latter system, does not require the design of a new dual-labeled detection oligonucleotide for each new target sequence.
Original languageEnglish
Pages (from-to)89 - 95
Number of pages7
JournalAnalytical Biochemistry
Volume422
Issue number2
DOIs
Publication statusPublished - 15 Mar 2012
Externally publishedYes

Keywords

  • Quantitative
  • Real time
  • Polymerase chain reaction
  • Multiplex

Cite this