Zhang, Chunsun and Xing, Da (2010) Microfluidic gradient PCR (MG-PCR): a new method for microfluidic DNA amplification. BIOMEDICAL MICRODEVICES, 12 (1). pp. 1-12. ISSN 1387-2176 (Print) 1572-8781 (Online)
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Official URL: http://www.springerlink.com/content/313r57q540l454...
This study develops a new microfluidic DNA amplification strategy for executing parallel DNA amplification in the microfluidic gradient polymerase chain reaction (MG-PCR) device. The developed temperature gradient microfluidic system is generated by using an innovative fin design. The device mainly consists of modular thermally conductive copper flake which is attached onto a finned aluminum heat sink with a small fan. In our microfluidic temperature gradient prototype, a non-linear temperature gradient is produced along the gradient direction. On the copper flake of length 45Â mm, width 40Â mm and thickness 4Â mm, the temperature gradient easily spans the range from 97 to 52Â°C. By making full use of the hot (90â97Â°C) and cold (60â70Â°C) regions on the temperature gradient device, the parallel, two-temperature MG-PCR amplification is feasible. As a demonstration, the MG-PCR from three parallel reactions of 112-bp Escherichia coli DNA fragment is performed in a continuous-flow format, in which the flow of the PCR reagent in the closed loop is induced by the buoyancy-driven nature convection. Although the prototype is not optimized, the MG-PCR amplification can be completed in less than 45Â min. However, the MG-PCR thermocycler presented herein can be further scaled-down, and thus the amplification times and reagent consumption can be further reduced. In addition, the currently developed temperature gradient technology can be applied onto other continuous-flow MG-PCR systems or used for other analytical purposes such as parallel and combination measurements, and fluorescent melting curve analysis.
|Subjects:||Biomedical Science > Nanoscale biological processes|
Biomedical Science > Nanobiotechnology
|Deposited By:||M T V|
|Deposited On:||12 Feb 2010 13:55|
|Last Modified:||12 Feb 2010 13:55|
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