Issue 10, 2012

Real time quantitative amplification detection on a microarray: towards high multiplex quantitative PCR

Abstract

Quantitative real-time polymerase chain reaction (qrtPCR) is widely used as a research and diagnostic tool. Notwithstanding its many powerful features, the method is limited in the degree of multiplexing to about 6 due to spectral overlap of the available fluorophores. A new method is presented that allows quantitative amplification detection at higher multiplexing by the integration of amplification in solution and monitoring via hybridization to a microarray in real-time. This method does not require any manipulation of the PCR product and runs in a single closed chamber. Employing labeled primers, one of the main challenges is to measure surface signals against a high fluorescence background from solution. A compact, confocal scanner is employed, based on miniaturized optics from DVD technology and combined with a flat thermocycler for simultaneous scanning and heating. The feasibility of this method is demonstrated in singleplex with an analytical sensitivity comparable to routine qrtPCR.

Graphical abstract: Real time quantitative amplification detection on a microarray: towards high multiplex quantitative PCR

Article information

Article type
Technical Note
Submitted
10 Aug 2011
Accepted
24 Feb 2012
First published
03 Apr 2012

Lab Chip, 2012,12, 1897-1902

Real time quantitative amplification detection on a microarray: towards high multiplex quantitative PCR

A. Pierik, M. Boamfa, M. van Zelst, D. Clout, H. Stapert, F. Dijksman, D. Broer and R. Wimberger-Friedl, Lab Chip, 2012, 12, 1897 DOI: 10.1039/C2LC20740K

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements