Novel fluorescence detection technique for non-contact temperature sensing in microchip PCR

Mondal, Sudip and Venkataraman, V (2007) Novel fluorescence detection technique for non-contact temperature sensing in microchip PCR. In: Journal of Biochemical and Biophysical Methods, 70 (5). pp. 773-777.

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Abstract

DNA amplification using Polymerase Chain Reaction (PCR) in a small volume is used in Lab-on-a-chip systems involving DNA manipulation. For few microliters of volume of liquid, it becomes difficult to measure and monitor the thermal profile accurately and reproducibly, which is an essential requirement for successful amplification. Conventional temperature sensors are either not biocompatible or too large and hence positioned away from the liquid leading to calibration errors. In this work we present a fluorescence based detection technique that is completely biocompatible and measures directly the liquid temperature. PCR is demonstrated in a 3 ILL silicon-glass microfabricated device using non-contact induction heating whose temperature is controlled using fluorescence feedback from SYBR green I dye molecules intercalated within sensor DNA. The performance is compared with temperature feedback using a thermocouple sensor. Melting curve followed by gel electrophoresis is used to confirm product specificity after the PCR cycles. (c) 2007 Elsevier B.V. All rights reserved.

Item Type:	Journal Article
Publication:	Journal of Biochemical and Biophysical Methods
Publisher:	Elsevier Science
Additional Information:	Copyright of this article belongs to Elsevier Science.
Department/Centre:	Division of Physical & Mathematical Sciences > Physics
Date Deposited:	29 Mar 2010 07:31
Last Modified:	19 Sep 2010 05:58
URI:	http://eprints.iisc.ac.in/id/eprint/26665

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