![]() doi: 10.1038/nmeth898ĭiehl F, Schmidt K, Durkee KH, Moore KJ, Goodman SN, Shuber AP, Kinzler KW, Vogelstein B (2008) Analysis of mutations in DNA isolated from plasma and stool of colorectal cancer patients. doi: 10.1073/pnas.0507904102ĭiehl F, Li M, He YP, Kinzler KW, Vogelstein B, Dressman D (2006) BEAMing: single-molecule PCR on microparticles in water-in-oil emulsions. doi: 10.1093/nar/ĭiehl F, Li M, Dressman D, He YP, Shen D, Szabo S, Diaz LA, Goodman SN, David KA, Juhl H, Kinzler KW, Vogelstein B (2005) Detection and quantification of mutations in the plasma of patients with colorectal tumors. doi: 10.1021/ac900060rīrownie J, Shawcross S, Theaker J, Whitcombe D, Ferrie R, Newton C, Little S (1997) The elimination of primer–dimer accumulation in PCR. doi: 10.1002/3īeyor N, Yi LN, Seo TS, Mathies RA (2009) Integrated capture, concentration, polymerase chain reaction, and capillary electrophoretic analysis of pathogens on a chip. doi: 10.1093/nar/28.20.e87īettiol S, Thompson MJ, Roberts NW, Perera R, Heneghan CJ, Harnden A (2010) Symptomatic treatment of the cough in whooping cough. These results show that this method is an effective method of DNA detection which is easily integrated in a microfluidic device to perform additional steps such as sample pre-conditioning.Īdessi C, Matton G, Ayala G, Turcatti G, Mermod J-J, Mayer P, Kawashima E (2000) Solid phase DNA amplification: characterisation of primer attachment and amplification mechanisms. We then demonstrate the use of the procedure in an integrated device capturing, amplifying, detecting, and purifying template DNA in a single microfluidic chamber. The device is capable of detecting a synthetically prepared section of the Bordetella pertussis genome in as few as 10 temperature cycles with times as short as 15 min. The effects of key bead-based PCR parameters, including annealing temperature and concentration of microbeads in the reaction mixture, are studied to achieve optimized device sensitivity and detection time. ![]() The microchip uses an integrated microheater and temperature sensor for rapid control of thermal cycling temperatures, while the sample is held in a microchamber fabricated from (poly)dimethylsiloxane and coated with Parylene. We leverage the ability of bead-based PCR to accumulate fluorescent labels following DNA amplification to explore a novel DNA detection scheme on a microchip. Bead-based PCR, while not extensively investigated in microchip format, has been used in a variety of bioanalytical applications in recent years. We present a bead-based approach to microfluidic polymerase chain reaction (PCR), enabling fluorescent detection and sample conditioning in a single microchamber.
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