Flow-through polymerase chain reactions in chip thermocyclers

Schneegaß, Ivonne; Köhler, J. Michael

doi:10.1016/s1389-0352(01)00033-2

Cited by 87 publications

(44 citation statements)

References 49 publications

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“…It was hypothesized that having a serpentine channel of varying widths could potentially cause flow recirculation that would nucleate bubbles in the regions with the expanding and contracting conduits [10]. Bubbles induce temperature gradients within the sample that significantly reduce or inhibit amplification while flow recirculation increases flow resistance (and may nucleate bubbles).…”

Section: Flow Simulations Within the Serpentine Microchannelmentioning

confidence: 99%

“…For the μ-PIV experiments, the image diameter for a particle in the object plane can be approximated as [27] ( 10) where M is the magnification,d p is the particle diameter, and d s is the diffraction-limited spot size. When d e = 1.9 µ m, the measured particle displacement in the μ-PIV experiments were accurate to within 0.19 µm.…”

Section: E μ-Piv Velocity Field Measurementsmentioning

confidence: 99%

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A Continuous-Flow Polymerase Chain Reaction Microchip With Regional Velocity Control

Fozdar

Ali

et al. 2006

J. Microelectromech. Syst.

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This paper presents a continuous-flow polymerase chain reaction (PCR) microchip with a serpentine microchannel of varying width for "regional velocity control." Varying the channel width by incorporating expanding and contracting conduits made it possible to control DNA sample velocities for the optimization of the exposure times of the sample to each temperature phase while minimizing the transitional periods during temperature transitions. A finite element analysis (FEA) and semianalytical heat transfer model was used to determine the distances between the three heating assemblies that are responsible for creating the denaturation (96 °C), hybridization (60 °C), and extension (72 °C) temperature zones within the microchip. Predictions from the thermal FEA and semi-analytical model were compared with temperature measurements obtained from an infrared (IR) camera. Flow-field FEAs were also performed to predict the velocity distributions in the regions of the expanding and contracting conduits to study the effects of the microchannel geometry on flow recirculation and bubble nucleation. The flow fields were empirically studied using micro particle image velocimetry (μ-PIV) to validate the flow-field FEA's and to determine experimental velocities in each of the regions of different width. Successful amplification of a 90 base pair (bp) bacillus anthracis DNA fragment was achieved.

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Section: Flow Simulations Within the Serpentine Microchannelmentioning

confidence: 99%

Section: E μ-Piv Velocity Field Measurementsmentioning

confidence: 99%

A Continuous-Flow Polymerase Chain Reaction Microchip With Regional Velocity Control

Fozdar

Ali

et al. 2006

J. Microelectromech. Syst.

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“…The development of microdevices for the Polymerase Chain Reaction (PCR) started in the early 1990s [41][42][43][44]. The reduction of PCR volume and the use of highly heat conducting materials as silicon allowed to reduce the times for thermocycling considerably [45].…”

Section: Pcr In Micro Fluid Segmentsmentioning

confidence: 99%

Droplet-Based Microfluidics as a Biomimetic Principle: From PCR-Based Virus Diagnostics to a General Concept for Handling of Biomolecular Information

Köhler

2012

Microdroplet Technology

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The ability of lithographic fabrication of microchannels for fluid transport on the one hand and the continuously decreasing critical dimensions during the development of microlithography over the last decades generated high expectations on the progress of automated processing of small amounts of substances. In particular, it was expected that large numbers of well distinguished chemical entities-atoms, molecules, nano particles-could become manipulated highly parallel and with high speed. This hope was fed by the imagination of an analogy between the electron transport in integrated circuits and the transport of chemical objects inside microfluidic networks. The experiences of the last both decades had shown that such ideas of a complete analogy are not realistic. The progress of handling of substance-coupled information is much slower as the progress of hard ware development in the electronic devices. But, beside this general disappointment, microfluidics is still promising the arise and growth-up of very important new strategies for organizing chemical systems at the microscale and for supplying functional interfaces between the complex information inside the world of molecules and particles on the one hand and electronic systems on the other hand.Whereas clouds of electrons are manipulated in digital electronic devices, clouds of molecules are transported through microchannels by a convective flow. But, the special conditions in microfluidics cause low Reynolds numbers which reflect the small ratio of channel diameter and volume flow rate to the viscous forces. The microfluidic conditions cause always a laminary structure of flow with steep flow velocity gradients and results in to very high fluidic dispersion of concentration signals if homogeneous fluids are applied. So, the principle high potential of J.M. K€ ohler

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“…An important trend in chemical and biological analysis over the past 15 years has been the miniaturization of analytical procedures and the development of micro-miniature analyzers (microchips) [1,2,3,4]. The ultimate goal of this work is a lab-on-a-chip or a µTAS (micro total analytical system) in which all the steps in an analytical procedure are performed in a single chip [5,6].…”

Section: Introductionmentioning

confidence: 99%

Microchip PCR

Kricka

Wilding

2003

Analytical and Bioanalytical Chemistry

145

107

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Miniaturization of genetic tests has become an important goal. This review surveys the current progress towards the miniaturization of tests based on the polymerase chain reaction (PCR). It examines the different types of PCR microchip designs, fabrication methods,and the components of a microchip PCR device. It also discusses the problems attributable to surface chemistry of microchip components (inhibition of PCR), and the static and dynamic surface passivation strategies developed for the solution of these difficulties

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Flow-through polymerase chain reactions in chip thermocyclers

Cited by 87 publications

References 49 publications

A Continuous-Flow Polymerase Chain Reaction Microchip With Regional Velocity Control

A Continuous-Flow Polymerase Chain Reaction Microchip With Regional Velocity Control

Droplet-Based Microfluidics as a Biomimetic Principle: From PCR-Based Virus Diagnostics to a General Concept for Handling of Biomolecular Information

Microchip PCR

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