Use of a Multiplexed CMOS Microarray to Optimize and Compare Oligonucleotide Binding to DNA Probes Synthesized or Immobilized on Individual Electrodes

Sensors

Karl Maurer²  Nina Yazvenko²  Jodi Wilmoth²  John Cooper²  Wanda Lyon¹ 

[1] AFRL/RHPB, Wright Patterson AFB, 2729 R Street, Area B B837, Dayton, OH 45433, USA; E-Mail:;CombiMatrix Corporation, 6500 Harbour Heights Parkway, Suite #202, Mukilteo WA 98275, USA; E-Mails:

关键词: CombiMatrix microarray;    biosensor;    electrochemical detection;    DNA;    hybridization;    polypyrrole;   

DOI  :  10.3390/s100807371

来源: mdpi

【 摘 要 】

The CombiMatrix microarray with 12,544 electrodes supports in situ electrochemical synthesis of user-defined DNA probes. As an alternative, we immobilized commercially synthesized DNA probes on individual electrodes coated with electropolymerized polypyrrole (Ppy). Hybridization was measured using a biotinylated target oligonucleotide and either Cy5-streptavidin and fluorescence detection or horseradish peroxidase-streptavidin and enzyme-enhanced electrochemical detection. Detection efficiencies were optimized by varying the deposition of the Ppy, the terminal groups on the DNA probes, and other factors that impacted fluorescence quenching and electrical conductivity. Optimized results were compared against those obtained using a microarray with the same DNA sequences synthesized in situ. Immobilized probes produced higher fluorescence signals, possibly by providing a greater stand off between the Cy5 on the target oligonucleotide and the quenching effects of the Ppy and the platinum electrode.

【 授权许可】

CC BY   
© 2010 by the authors; licensee MDPI, Basel, Switzerland.