Influence of segmenting fluids on efficiency, crossing point and fluorescence level in real time quantitative PCR

Edmond J. Walsh, C. King, R. Grimes, A. Gonzalez

Research output: Contribution to journalArticlepeer-review

Abstract

The two-phase segmented flow approach to the processing and quantitative analysis of biological samples in microdevices offers significant advantages over the single-phase continuous flow methodology. Despite this, little is known about the compatibility of samples and reactants with segmenting fluids, although a number of investigators have reported reduced yield and inhibition of enzymatic reactions depending on the segmenting fluid employed. The current study addresses the compatibility of various segmenting fluids with real time quantitative PCR to understand the physicochemical requirements of this important reaction in biotechnology. The results demonstrate that creating a static segmenting fluid/PCR mix interface has a negligible impact on the reaction efficiency, crossing threshold and end fluorescence levels using a variety of segmenting fluids. The implication is then that the previously reported inhibitory effects are the result of the dynamic motion between the segmenting fluid and the sample in continuously flowing systems. The results presented here are a first step towards understanding the limitations of the segmented flow methodology, which are necessary to bring this approach into mainstream use.

Original languageEnglish
Pages (from-to)59-64
Number of pages6
JournalBiomedical microdevices
Volume8
Issue number1
DOIs
Publication statusPublished - Mar 2006

Keywords

  • Biocompatible materials
  • Microfluidic analytical techniques
  • Polymerase chain reaction

Fingerprint

Dive into the research topics of 'Influence of segmenting fluids on efficiency, crossing point and fluorescence level in real time quantitative PCR'. Together they form a unique fingerprint.

Cite this