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Merck

Rapid DNA amplification using a battery-powered thin-film resistive thermocycler.

Methods in molecular biology (Clifton, N.J.) (2009-01-23)
Keith E Herold, Nikolay Sergeev, Andriy Matviyenko, Avraham Rasooly
RESUMEN

A prototype handheld, compact, rapid thermocycler was developed for multiplex analysis of nucleic acids in an inexpensive, portable configuration. Instead of the commonly used Peltier heating/cooling element, electric thin-film resistive heater and a miniature fan enable rapid heating and cooling of glass capillaries leading to a simple, low-cost Thin-Film Resistive Thermocycler (TFRT). Computer-based pulse width modulation control yields heating rates of 6-7 K/s and cooling rates of 5 K/s. The four capillaries are closely coupled to the heater, resulting in low power consumption. The energy required by a nominal PCR cycle (20 s at each temperature) was found to be 57+/-2 J yielding an average power of approximately 1.0 W (not including the computer and the control system). Thus the device can be powered by a standard 9 V alkaline battery (or other 9 V power supply). The prototype TFRT was demonstrated (in a benchtop configuration) for detection of three important food pathogens (E. coli ETEC, Shigella dysenteriae, and Salmonella enterica). PCR amplicons were analyzed by gel electrophoresis. The 35 cycle PCR protocol using a single channel was completed in less then 18 min. Simple and efficient heating/cooling, low cost, rapid amplification, and low power consumption make the device suitable for portable DNA amplification applications including clinical point of care diagnostics and field use.

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Sigma-Aldrich
Seroalbúmina bovina, lyophilized powder, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Ethidium bromide solution, BioReagent, for molecular biology, 10 mg/mL in H2O
Sigma-Aldrich
ADN polimerasa Taq from Thermus aquaticus, with 10× PCR reaction buffer without MgCl2
Sigma-Aldrich
ADN polimerasa Taq from Thermus aquaticus, with 10× PCR reaction buffer containing MgCl2
Sigma-Aldrich
Aceite de inmersión, PCR Reagent