Design and Development of Convective Flow Apparatus for Pcr Thermocycling

Jennifer L. Christensen, Chemical Engineering, Texas A&M University, PO Box 13804, College Station, TX 77841 and Victor M. Ugaz, Artie McFerrin Department of Chemical Engineering, Texas A&M University, 3122 TAMU, College Station, TX 77843.

The Polymerase Chain Reaction (PCR) is the productive process that duplicates targeted DNA strands based on a varying temperature cycle. The use of the PCR played a crucial part in the Human Genome Project, continues to be an integral part of studying disease, and is earning its role in the world of forensics. Currently, PCR faces a time issue, as it is typically ran in a thermocycler, averaging between one and three hours. Using a convective flow system, the process runs faster and more efficient, using natural convection and buoyancy forces to create the required temperature cycles. The objective was to redesign the convective flow setup by using a heater on top and bottom calibrated and independently controlled, as well as a clear, multi-well plexiglass block to hold 30 uL of sample per well. The device has produced inconsistent results, mainly due to the unpredictable nature of varying reactions used; however, the ultimate goal of reduced time requirements has been met. To test the sensitivity and efficiency of a reaction, a dilution of the template DNA was prepared and ran in the thermocycler to find the minimum number of detectable copies. Therefore, using the thermocycler as a control for the convective flow experiment, we are effectively testing many reactions in order to isolate a workable reaction and optimize the convective flow parameters.