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Label-free electrical detection of pyrophosphate generated from DNA polymerase reactions on field-effect devices.


ABSTRACT: We introduce a label-free approach for sensing polymerase reactions on deoxyribonucleic acid (DNA) using a chelator-modified silicon-on-insulator field-effect transistor (SOI-FET) that exhibits selective and reversible electrical response to pyrophosphate anions. The chemical modification of the sensor surface was designed to include rolling-circle amplification (RCA) DNA colonies for locally enhanced pyrophosphate (PPi) signal generation and sensors with immobilized chelators for capture and surface-sensitive detection of diffusible reaction by-products. While detecting arrays of enzymatic base incorporation reactions is typically accomplished using optical fluorescence or chemiluminescence techniques, our results suggest that it is possible to develop scalable and portable PPi-specific sensors and platforms for broad biomedical applications such as DNA sequencing and microbe detection using surface-sensitive electrical readout techniques.

SUBMITTER: Credo GM 

PROVIDER: S-EPMC3367324 | biostudies-literature | 2012 Mar

REPOSITORIES: biostudies-literature

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Label-free electrical detection of pyrophosphate generated from DNA polymerase reactions on field-effect devices.

Credo Grace M GM   Su Xing X   Wu Kai K   Elibol Oguz H OH   Liu David J DJ   Reddy Bobby B   Tsai Ta-Wei TW   Dorvel Brian R BR   Daniels Jonathan S JS   Bashir Rashid R   Varma Madoo M  

The Analyst 20120119 6


We introduce a label-free approach for sensing polymerase reactions on deoxyribonucleic acid (DNA) using a chelator-modified silicon-on-insulator field-effect transistor (SOI-FET) that exhibits selective and reversible electrical response to pyrophosphate anions. The chemical modification of the sensor surface was designed to include rolling-circle amplification (RCA) DNA colonies for locally enhanced pyrophosphate (PPi) signal generation and sensors with immobilized chelators for capture and su  ...[more]

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