The proposed research investigates the hitherto unknown fundamental characteristics of electrostatic DNA hybridization. Namely; the yield, kinetic rates, ability to denature long DNA and to support a sufficient number of hybridization cycles, in order to establish conditions for efficient PCR amplification.
The specific aims are to:
1. Construct a fluorescent detector for hybridization and extension of electrode tethered primers. An evanescent wave fluorescent real-time detector will be built to measure the kinetics and yield of denaturation, annealing, and extension stages in solid-phase PCR.
2. Prove-in-principle and optimize electrostatic isothermal PCR. Linear and exponential amplification will be demonstrated in real-time PCR mode using oligonucleotide templates. Efficient electrostatic pulse cycles and optimal surface and solution conditions will be defined. Amplification protocols will be developed.