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Dynamic optimization of on-chip polymerase chain reaction by monitoring intracycle fluorescence using fast synchronous detection

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3 Author(s)
Mondal, Sudip ; Department of Physics, Indian Institute of Science, Bangalore 560 012, India ; Paul, Debjani ; Venkataraman, V.

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The authors report on-chip dynamic optimization of polymerase chain reaction (PCR) based on a feedback technique utilizing synchronous detection of intracycle fluorescence every 500 ms. From a direct measurement of polymerase activity, the authors determine the optimum extension temperature. The authors dynamically optimize PCR in an inductively heated microchip by sensing the saturation of extension in each cycle and applying the feedback. They demonstrate that, even with fast ramp rates, dynamic optimization leads to faster reactions compared to fixed-duration extension protocols for long DNA (≫500 bp). This optimization scheme uses a fairly universal dye Sybr Green I and can be applied to most PCRs.

Published in:

Applied Physics Letters  (Volume:90 ,  Issue: 1 )

Date of Publication:

Jan 2007

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