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

Analyst. 2012 Mar 21;137(6):1351-62. doi: 10.1039/c2an15930a. Epub 2012 Jan 19.

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.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Base Sequence
  • Biosensing Techniques
  • Chelating Agents / chemistry
  • DNA / chemistry*
  • DNA / metabolism*
  • DNA-Directed DNA Polymerase / chemistry
  • DNA-Directed DNA Polymerase / metabolism*
  • Diphosphates / analysis*
  • Electrochemical Techniques*
  • Molecular Sequence Data
  • Molecular Structure
  • Silanes / chemistry
  • Silicon / chemistry
  • Surface Properties
  • Transistors, Electronic*

Substances

  • Chelating Agents
  • Diphosphates
  • Silanes
  • DNA
  • DNA-Directed DNA Polymerase
  • Silicon