Reactions and Fluidics in Miniaturized Natural Convection Systems

Buoyancy-driven convection offers a novel and greatly simplified mechanism for generating continuous nonpulsatile flow fields and performing thermally activated biochemical reactions. In this paper, we build on our previous work by constructing a multiwell device incorporating an array of 35-μL cyli...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2004-11, Vol.76 (21), p.6254-6265
Main Authors: Krishnan, Madhavi, Agrawal, Nitin, Burns, Mark A, Ugaz, Victor M
Format: Article
Language:English
Subjects:
Actins - chemistry
Analytical chemistry
Base Sequence
Biological and medical sciences
Biotechnology
Buoyancy
Chemical reactions
DNA Primers
Fluids
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Humans
In vitro gene amplification. Pcr technique
Methods. Procedures. Technologies
Miniaturization
Polymerase Chain Reaction
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Summary:Buoyancy-driven convection offers a novel and greatly simplified mechanism for generating continuous nonpulsatile flow fields and performing thermally activated biochemical reactions. In this paper, we build on our previous work by constructing a multiwell device incorporating an array of 35-μL cylindrical cavities to perform polymerase chain reaction (PCR) amplification of a 191-base pair fragment associated with membrane channel proteins M1 and M2 of the influenza-A virus in as little as 15 min with performance comparable to conventional thermocyclers. We also describe entirely new adaptations of convective flows by conducting a series of coordinated flow visualization and computational studies to explore the design of closed-loop systems to execute tunable thermocycling, pumping, and mixing operations in a format suitable for integration into miniaturized biochemical analysis systems. Using 15-μL convective flow loops, we are able to perform PCR amplification of the same 191-base pair fragment associated with the influenza-A virus, as well as a 295-base pair segment of the human β-actin gene in a format offering an enhanced degree of control and tunability. These convective flow devices can be further scaled down to nanoliter volumes and are ideally suited as a platform for a new generation of low-power, portable microfluidic DNA analysis systems.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac049323u