Rapid microfluidic thermal cycler for polymerase chain reaction nucleic acid amplification

Mahjoob, Shadi; Vafai, Kambiz; Beer, Neil Reginald

doi:10.1016/j.ijheatmasstransfer.2007.11.014

Cited by 33 publications

(13 citation statements)

References 42 publications

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“…Indeed, the regulation of temperature is a critical parameter in managing many physical, chemical and biological applications. Prominent examples of applications requiring tight temperature control are Polymerase Chain Reaction (PCR) [ 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 ], Temperature Gradient Focusing for Electrophoresis (TGF) [ 16 , 17 ], digital microfluidics [ 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 ], mixing [ 32 , 33 , 34 ], and protein crystallization [ 35 ]. The scope of this article is to provide a comprehensive applications-based overview of heating techniques reported in the literature during the last decade.…”

Section: Introductionmentioning

confidence: 99%

A Review of Heating and Temperature Control in Microfluidic Systems: Techniques and Applications

et al. 2013

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This review presents an overview of the different techniques developed over the last decade to regulate the temperature within microfluidic systems. A variety of different approaches has been adopted, from external heating sources to Joule heating, microwaves or the use of lasers to cite just a few examples. The scope of the technical solutions developed to date is impressive and encompasses for instance temperature ramp rates ranging from 0.1 to 2,000 °C/s leading to homogeneous temperatures from −3 °C to 120 °C, and constant gradients from 6 to 40 °C/mm with a fair degree of accuracy. We also examine some recent strategies developed for applications such as digital microfluidics, where integration of a heating source to generate a temperature gradient offers control of a key parameter, without necessarily requiring great accuracy. Conversely, Temperature Gradient Focusing requires high accuracy in order to control both the concentration and separation of charged species. In addition, the Polymerase Chain Reaction requires both accuracy (homogeneous temperature) and integration to carry out demanding heating cycles. The spectrum of applications requiring temperature regulation is growing rapidly with increasingly important implications for the physical, chemical and biotechnological sectors, depending on the relevant heating technique.

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Section: Introductionmentioning

confidence: 99%

A Review of Heating and Temperature Control in Microfluidic Systems: Techniques and Applications

et al. 2013

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“…These advantages have been utilized over the past several years for purposes of amplifying nucleic acids on chips (polymerase chain reaction, PCR) (Ke et al 2006;Liao et al 2005). Mahjoob et al (2008) have proposed the insertion of a porous matrix to increase heating/cooling rates due a very large surface area for a given volume, which is a key parameter in heat transfer processes. Although, elegant and efficient, this approach requires a precise control of many parameters, such as exchanger geometry, flow rate, conductive plate, and the porous matrix material, which decrease the flexibility of this method.…”

Section: Introductionmentioning

confidence: 99%

Integration of a uniform and rapid heating source into microfluidic systems

Selva

Mary

Jullien

2009

Microfluid Nanofluid

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The purpose of this study has been to demonstrate the possibilities of uniform heating of a cavity, with great accuracy, by means of an integrated resistor built with the same dimensions as the cavity, i.e., with a high level of integration. For application purposes, a compact resistor allows increasing the number of cavities in which temperature can be independently controlled on the same substrate, which can prove critical for high-throughput screening applications. Potential applications lie in the field of biology or chemistry. In order to achieve the desired result, an optimization procedure was performed on the shape of the resistor. The heater size reduction enables a high level of integration with a reduced heating source surface area. Resistor shape has been optimized to reduce the influence of boundary effects, using improvements introduced in genetic algorithms. An experimental validation of the temperature profile inside the cavity has been carried out using a dye whose fluorescence depends on temperature, i.e., Rhodamine B, it will be shown herein that the optimized resistor allows for temperature cycling, e.g., for PCR applications.

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“…Although a field-portable realtime PCR machine is available (Idaho Technology, Salt Lake City, UT, USA), it is expensive, heavy, and regulated by the United States government. Miniaturization of PCR technology for point-of-use application is an active area of research (Mahjoob et al 2008;Maltezos et al 2008;Xiang et al 2007), but existing systems remain impractical for field use.…”

Section: Introductionmentioning

confidence: 99%

In silico genomic subtraction guides development of highly accurate, DNA-based diagnostics for Ralstonia solanacearum race 3 biovar 2 and blood disease bacterium

et al. 2011

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New rapid diagnostic methods are urgently needed to discriminate the quarantine pathogen Ralstonia solanacearum (Rs) race 3 biovar 2 (R3B2) from other populations of Rs that lack the adaptation to cause bacterial wilt disease in temperate regions. We used an in silico bioinformatic approach to identify several genome sequences potentially specific to R3B2 strains. Primer sets were designed to PCR-amplify sequences in these regions, and four sets were ultimately shown to be [99% accurate for detection of R3B2 strains. On the basis of these results, several primers were designed to enable development of a loop-mediated isothermal amplification assay that was rapid, technologically simple, and essentially 100% accurate for identification of R3B2 when applied to a comprehensive collection of geographically diverse Rs strains. We fortuitously found that a sequence in one of the ''R3B2-specific'' regions has *90% identity to a sequence present in strains of the blood disease bacterium (BDB), a member of the Rs species complex that infects banana. Alignments of these sequences allowed design of a second PCR primer set that proved 100% accurate for identification of BDB strains when tested on the 22 BDB strains available to us. These results demonstrate the power of in silico genomic subtraction for rapid identification of population-specific DNA sequences and for the development of simple, reliable detection methods for Rs subpopulations.

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Rapid microfluidic thermal cycler for polymerase chain reaction nucleic acid amplification

Cited by 33 publications

References 42 publications

A Review of Heating and Temperature Control in Microfluidic Systems: Techniques and Applications

A Review of Heating and Temperature Control in Microfluidic Systems: Techniques and Applications

Integration of a uniform and rapid heating source into microfluidic systems

In silico genomic subtraction guides development of highly accurate, DNA-based diagnostics for Ralstonia solanacearum race 3 biovar 2 and blood disease bacterium

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