Single-molecule reverse transcription polymerase chain reaction using water-in-oil emulsion

J Biosci Bioeng. 2005 Mar;99(3):293-5. doi: 10.1263/jbb.99.293.

Abstract

A new method based on a combination of reverse transcription polymerase chain reaction (RT-PCR) and a water-in-oil (W/O) emulsion was developed. Reverse transcription and initial thermal cycles were carried out in droplets of the W/O emulsion. Then, the droplets were united, followed by remaining secondary PCR cycles. This two-step method succeeded in detecting a single RNA molecule.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chemical Fractionation / methods
  • Emulsions / chemistry
  • HIV / genetics*
  • HIV / isolation & purification*
  • Oils / chemistry*
  • RNA, Viral / analysis*
  • RNA, Viral / chemistry
  • RNA, Viral / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Water / chemistry*

Substances

  • Emulsions
  • Oils
  • RNA, Viral
  • Water