Label-free microelectronic PCR quantification.
We present a robust and simple method for direct, label-free PCR product quantification using an integrated microelectronic sensor. The field-effect sensor can sequentially detect the intrinsic charge of multiple unprocessed PCR products and does not require sample processing or additional reagents in the PCR mixture. The sensor measures nucleic acid concentration in the PCR relevant range and specifically detects the PCR products over reagents such as Taq polymerase and nucleotide monomers. The sensor can monitor the product concentration at various stages of PCR and can generate a readout that resembles that of a real-time fluorescent measurement using an intercalating dye but without its potential inhibition artifacts. The device is mass-produced using standard semiconductor processes, can be reused for months, and integrates all sensing components directly on-chip. As such, our approach establishes a foundation for the direct integration of PCR-based in vitro biotechnologies with microelectronics.
Hou
Chih-Sheng Johnson
CS
Department of Electrical Engineering and Computer Science, Biological Engineering Division, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Milovic
Nebojsa
N
Godin
Michel
M
Russo
Peter R
PR
Chakrabarti
Raj
R
Manalis
Scott R
SR
D004581:Electronics; D005456:Fluorescent Dyes / Q000737:chemistry*; D007364:Intercalating Agents / Q000737:chemistry*; D008832:Microchemistry / Q000295:instrumentation / Q000379:methods*; D009696:Nucleic Acids / Q000032:analysis*; D009711:Nucleotides / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D015203:Reproducibility of Results; D012666:Semiconductors; D012680:Sensitivity and Specificity; D019914:Taq Polymerase / Q000737:chemistry
D005456:Fluorescent Dyes; D007364:Intercalating Agents; D009696:Nucleic Acids; D009711:Nucleotides; D019914:Taq Polymerase