Abstract
Rapid detection of target DNA sequences in complex matrices is possible through the obviation of pre-PCR target ssDNA and post-PCR amplicon isolation steps. We demonstrate the sensing abilities of single-strand (ss)DNA tethered to GUITAR electrodes to recognize targets by hybridization. The formation of the corresponding duplex DNA is detected with a voltammetric signal with surface-bound cobalt (III) phenanthroline complex. In cases where the capture of target ssDNA does not fully saturate the surface bound complementary sequences, we employ rapid PCR (17 mins) directly on the electrode. The first part of this dissertation would introduce the concept of on-Electrode (oE) PCR protocol. This involves the use of modified GUITAR coated on a carbon fiber to immobilize ssDNA primers for the rapid detection of Salmonella ssDNA through the oE-PCR protocol. The next chapter of this dissertation will explore application of oE-PCR to target Nitrobacter genomic double stranded (ds)DNA in a wastewater matrix. The third chapter of this dissertation would investigate the application of GUITAR PCR electrodes to target chicken genomic dsDNA. The final part of this dissertation will explore a new functionalization pathway for GUITAR electrode with DNA that would be applied in the ultrasensitive detection of silver ions in water. This new technique demonstrated will allow investigators to make rapid field observations of disease detection and rapid heavy metal analysis. Future applications will include making this into a chip for medical and forensic rapid DNA diagnostics..