Convectively Driven Polymerase Chain Reaction Thermal Cycler

Wheeler, Elizabeth K.; Benett, William J.; Stratton, Paul; Richards, James B.; Chen, A.; Christian, Allen T.; Ness, Kevin; Ortega, Jason; Li, L. G.; Weisgraber, Todd H.; Goodson, Kenneth E.; Milanovich, Fred P.

doi:10.1021/ac034941g

Cited by 102 publications

(81 citation statements)

References 11 publications

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“…In certain applications such as magneto-hydrodynamically (MHD) driven circular chromatography, 21 MHD-driven PCR, 22 MHD stirrer, 23 and self-actuated flow-cycling PCR, 24,25 it is desirable to flow reagents in a closed loop. In a pneumatic system, the filling of the closed loop without creating gas bubbles represents a challenge.…”

Section: Filling and Withdrawing Samples From A Closed Loopmentioning

confidence: 99%

Thermally-actuated, phase change flow control for microfluidic systems

et al. 2005

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An easy to implement, thermally-actuated, noninvasive method for flow control in microfluidic devices is described. This technique takes advantage of the phase change of the working liquid itself-the freezing and melting of a portion of a liquid slug-to noninvasively close and open flow passages (referred to as a phase change valve). The valve was designed for use in a miniature diagnostic system for detecting pathogens in oral fluids at the point of care. The paper describes the modeling, construction, and characteristics of the valve. The experimental results favorably agree with theoretical predictions. In addition, the paper demonstrates the use of the phase change valves for flow control, sample metering and distribution into multiple analysis paths, sealing of a polymerase chain reaction (PCR) chamber, and sample introduction into and withdrawal from a closed loop. The phase change valve is electronically addressable, does not require any moving parts, introduces only minimal dead volume, is leakage and contamination free, and is biocompatible. CommentsFor personal or professional use only; May not be further made available or distributed.

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Section: Filling and Withdrawing Samples From A Closed Loopmentioning

confidence: 99%

Thermally-actuated, phase change flow control for microfluidic systems

et al. 2005

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“…In 2004, Wheeler et al developed a convectively driven, silicon-based PCR thermal chamber that utilized buoyancy forces to move the reaction fluid between certain temperatures required for successful PCR amplification. [56] The connectively driven polymerase chain reaction (CPCR) device differs from previous flow-through devices by eliminating the need of external pumping mechanisms due to the closed loop design of the CPCR system. [56] This CF format device also achieves a more efficient PCR reaction by the geometry of the heaters on the sides, instead of the top and bottom, of the device.…”

Section: Microfluidicsmentioning

confidence: 99%

“…[56] The connectively driven polymerase chain reaction (CPCR) device differs from previous flow-through devices by eliminating the need of external pumping mechanisms due to the closed loop design of the CPCR system. [56] This CF format device also achieves a more efficient PCR reaction by the geometry of the heaters on the sides, instead of the top and bottom, of the device. [56] Thus, miniaturization allows integration of all functional steps of DNA analysis into a single microchip device.…”

Section: Microfluidicsmentioning

confidence: 99%

Flow through PCR module of BioBriefcase

et al. 2005

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The BioBriefcase is an integrated briefcase-sized aerosol collection and analysis system for autonomous monitoring of the environment, which is currently being jointly developed by Lawrence Livermore and Sandia National Laboratories. This poster presents results from the polymerase chain reaction (PCR) module of the system. The DNA must be purified after exiting the aerosol collector to prevent inhibition of the enzymatic reaction. Traditional solid-phase extraction results in a large loss of sample. In this flow-through system, we perform sample purification, concentration and amplification in one reactor, which minimizes the loss of material. The sample from the aerosol collector is mixed with a denaturation solution prior to flowing through a capillary packed with silica beads. The DNA adheres to the silica beads allowing the environmental contaminants to be flushed to waste while effectively concentrating the DNA on the silica matrix. The adhered DNA is amplified while on the surface of the silica beads, resulting in a lower limit of detection than an equivalent eluted sample. Thus, this system is beneficial since more DNA is available for amplification, less reagents are utilized, and contamination risks are reduced.

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“…When the system reaches thermal equilibrium, the steady flow fields are developed and the reagents are ready to amplify DNA fragments in the tube. Although ccPCR is simple and easy-to-use, the efficiency of amplification could be varied because solutions could circulate around either the inner or outer paths [12][13][14][15]. This consequence is unfavorable for the quantification of copy numbers.…”

Section: Introductionmentioning

confidence: 99%

Development of Capillary Loop Convective Polymerase Chain Reaction Platform with Real-Time Fluorescence Detection

et al. 2017

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Polymerase chain reaction (PCR) has been one of the principal techniques of molecular biology and diagnosis for decades. Conventional PCR platforms, which work by rapidly heating and cooling the whole vessel, need complicated hardware designs, and cause energy waste and high cost. On the other hand, partial heating on the various locations of vessels to induce convective solution flows by buoyancy have been used for DNA amplification in recent years. In this research, we develop a new convective PCR platform, capillary loop convective polymerase chain reaction (clcPCR), which can generate one direction flow and make the PCR reaction more stable. The U-shaped loop capillaries with 1.6 mm inner diameter are designed as PCR reagent containers. The clcPCR platform utilizes one isothermal heater for heating the bottom of the loop capillary and a CCD device for detecting real-time amplifying fluorescence signals. The stable flow was generated in the U-shaped container and the amplification process could be finished in 25 min. Our experiments with different initial concentrations of DNA templates demonstrate that clcPCR can be applied for precise quantification. Multiple sample testing and real-time quantification will be achieved in future studies.

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Convectively Driven Polymerase Chain Reaction Thermal Cycler

Cited by 102 publications

References 11 publications

Thermally-actuated, phase change flow control for microfluidic systems

Thermally-actuated, phase change flow control for microfluidic systems

Flow through PCR module of BioBriefcase

Development of Capillary Loop Convective Polymerase Chain Reaction Platform with Real-Time Fluorescence Detection

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