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Ultrasensitive PCR and real-time detection from human genomic samples using a bidirectional flow microreactor

Ultrasensitive PCR and real-time detection from human genomic samples using a bidirectional flow microreactor
Ultrasensitive PCR and real-time detection from human genomic samples using a bidirectional flow microreactor
In this paper we present a reliable bidirectional flow DNA amplification microreactor for processing real-world genomic samples. This system shares the low-power thermal responsiveness of a continuous flow reactor with the low surface area to volume ratio character of stationary reactors for reducing surface inhibitory effects. Silanization with dimethyldichlorosilane in combination with dynamic surface passivation was used to enhance PCR compatibility and enable efficient amplification. For realtime fragment amplification monitoring we have implemented an epimodal fluorescent detection capability. The passivated bidirectional flow system was ultrasensitive, achieving an RNase P gene detection limit of 24 human genome copies with a reaction efficiency of 77%. This starts to rival the performance of a conventional real-time PCR instrument with a reaction efficiency of 93% and revitalizes flow-through PCR as a viable component of lab on a chip DNA analysis formats.
polymerase-chain-reaction nucleic-acid analysis dna analysis device capillary electrophoresis reaction amplification glass microchip chip cell chambers microfluidics
0003-2700
9185-9190
Chen, L.
586c0d55-dc72-49c0-a4fa-31df7000ce18
West, J.
f1c2e060-16c3-44c0-af70-242a1c58b968
Auroux, P. A.
18dc1e2e-6c2f-45fd-ba98-1a55e5e605cd
Manz, A.
d8d789e5-7066-4583-81f2-9d0410d3bbdc
Day, P. J. R.
b8ad0572-e75d-48bf-a25f-dfc9b1ed7b6a
Chen, L.
586c0d55-dc72-49c0-a4fa-31df7000ce18
West, J.
f1c2e060-16c3-44c0-af70-242a1c58b968
Auroux, P. A.
18dc1e2e-6c2f-45fd-ba98-1a55e5e605cd
Manz, A.
d8d789e5-7066-4583-81f2-9d0410d3bbdc
Day, P. J. R.
b8ad0572-e75d-48bf-a25f-dfc9b1ed7b6a

Chen, L., West, J., Auroux, P. A., Manz, A. and Day, P. J. R. (2007) Ultrasensitive PCR and real-time detection from human genomic samples using a bidirectional flow microreactor. Analytical Chemistry, 79 (23), 9185-9190. (doi:10.1021/ac701668k).

Record type: Article

Abstract

In this paper we present a reliable bidirectional flow DNA amplification microreactor for processing real-world genomic samples. This system shares the low-power thermal responsiveness of a continuous flow reactor with the low surface area to volume ratio character of stationary reactors for reducing surface inhibitory effects. Silanization with dimethyldichlorosilane in combination with dynamic surface passivation was used to enhance PCR compatibility and enable efficient amplification. For realtime fragment amplification monitoring we have implemented an epimodal fluorescent detection capability. The passivated bidirectional flow system was ultrasensitive, achieving an RNase P gene detection limit of 24 human genome copies with a reaction efficiency of 77%. This starts to rival the performance of a conventional real-time PCR instrument with a reaction efficiency of 93% and revitalizes flow-through PCR as a viable component of lab on a chip DNA analysis formats.

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More information

Published date: 2007
Additional Information: ISI Document Delivery No.: 236OH Times Cited: 20 Cited Reference Count: 47 Chen, Lin West, Jonathan Auroux, Pierre-Alain Manz, Andreas Day, Philip J. R. Amer chemical soc Washington
Keywords: polymerase-chain-reaction nucleic-acid analysis dna analysis device capillary electrophoresis reaction amplification glass microchip chip cell chambers microfluidics
Organisations: Cancer Sciences

Identifiers

Local EPrints ID: 346432
URI: http://eprints.soton.ac.uk/id/eprint/346432
ISSN: 0003-2700
PURE UUID: 344480cb-b35d-43ec-87f4-e5e8a934b78b
ORCID for J. West: ORCID iD orcid.org/0000-0002-5709-6790

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Date deposited: 05 Feb 2013 11:09
Last modified: 15 Mar 2024 03:43

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Contributors

Author: L. Chen
Author: J. West ORCID iD
Author: P. A. Auroux
Author: A. Manz
Author: P. J. R. Day

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