Issue 8, 2011

Nanolitre droplet array for real time reverse transcription polymerase chain reaction

Abstract

Recently, more and more effort has been put into the miniaturization of genetic tests such as quantitative PCR (qPCR), because it is no doubt a powerful tool for molecular diagnosis and quantitative biology. In this paper, we developed a low density nanolitre droplet array generated on a chemical modified silicon chip for gene quantification. Reliable and sensitive two step real time qRT-PCR assay for microRNA measurement was performed within 500 nL droplets. It has a dynamic range of six orders of magnitude, allowing for the quantification of microRNA input from 103 to 109 copies per reaction. We successfully applied the platform for quantitative measurement of mir-122 across five cultured cell lines. The minimum total RNA input was as low as 1 pg per assay, which showed great potential for gene quantification at single cell level. We envision the droplet based qPCR chip would be a universal and low-cost platform for gene quantification in ordinary biological laboratories.

Graphical abstract: Nanolitre droplet array for real time reverse transcription polymerase chain reaction

Supplementary files

Article information

Article type
Technical Note
Submitted
14 Oct 2010
Accepted
07 Feb 2011
First published
28 Feb 2011

Lab Chip, 2011,11, 1545-1549

Nanolitre droplet array for real time reverse transcription polymerase chain reaction

Y. Zhang, Y. Zhu, B. Yao and Q. Fang, Lab Chip, 2011, 11, 1545 DOI: 10.1039/C0LC00502A

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