Nucleic acid amplification of individual molecules in a microfluidic device.

@article{citeulike:3017896, abstract = {A microfluidic device was developed that enabled rapid polymerase chain reaction (PCR) analysis of individual DNA molecules. The device combined a means for accessing samples serially from a microtiter plate, channels for assembling eight parallel PCR reactions, and integrated resistive heaters for rapid thermocycling (>5 degrees C/s heating, >7 degrees C/s cooling) of samples as they flowed continuously through PCR channels. Amplification was monitored by fluorescence detection of Taqman probes. The long, narrow channels (10 microm x 180 microm x 40 mm) allowed sufficient separation between neighboring DNA templates to enable amplification of discreet DNA molecules. The functionality of the device was demonstrated by reproducibly amplifying a 2D6.6 CYP450 template and distinguishing between wild-type and mutant sequences using Taqman probes. A comparison of the rate of individual amplification events to the expected Poisson distribution confirmed that the device could reliably analyze individual DNA molecules. This work establishes the feasibility of rapid, single-molecule interrogation of nucleic acids.}, address = {Caliper Life Sciences, 605 Fairchild Drive, Mountain View, California 94024, USA.}, author = {Dettloff, R. and Yang, E. and Rulison, A. and Chow, A. and Farinas, J. }, citeulike-article-id = {3017896}, doi = {10.1021/ac800339w}, issn = {1520-6882}, journal = {Analytical chemistry}, keywords = {microfluidics}, month = {June}, number = {11}, pages = {4208--4213}, posted-at = {2008-07-18 14:12:16}, priority = {2}, title = {Nucleic acid amplification of individual molecules in a microfluidic device.}, url = {http://dx.doi.org/10.1021/ac800339w}, volume = {80}, year = {2008} }

TY - JOUR ID - citeulike:3017896 L3 - citeulike-article-id:3017896 TI - Nucleic acid amplification of individual molecules in a microfluidic device. JF - Analytical chemistry VL - 80 IS - 11 SP - 4208 EP - 4213 AD - Caliper Life Sciences, 605 Fairchild Drive, Mountain View, California 94024, USA. SN - 1520-6882 N2 - A microfluidic device was developed that enabled rapid polymerase chain reaction (PCR) analysis of individual DNA molecules. The device combined a means for accessing samples serially from a microtiter plate, channels for assembling eight parallel PCR reactions, and integrated resistive heaters for rapid thermocycling (>5 degrees C/s heating, >7 degrees C/s cooling) of samples as they flowed continuously through PCR channels. Amplification was monitored by fluorescence detection of Taqman probes. The long, narrow channels (10 microm x 180 microm x 40 mm) allowed sufficient separation between neighboring DNA templates to enable amplification of discreet DNA molecules. The functionality of the device was demonstrated by reproducibly amplifying a 2D6.6 CYP450 template and distinguishing between wild-type and mutant sequences using Taqman probes. A comparison of the rate of individual amplification events to the expected Poisson distribution confirmed that the device could reliably analyze individual DNA molecules. This work establishes the feasibility of rapid, single-molecule interrogation of nucleic acids. KW - microfluidics AU - Dettloff, R AU - Yang, E AU - Rulison, A AU - Chow, A AU - Farinas, J PY - 2008/06/01/ UR - http://dx.doi.org/10.1021/ac800339w ER -