Robert L. Brumley, Jr.
GeneSys Technologies, Inc., Mazomanie, WI 53560

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The completion of the Human Genome Project and other large scale projects require the development of new methodologies to significantly increase the throughput and decrease the cost of DNA mapping and sequencing. Currently, most DNA laboratories utilize several automated workstations operated in parallel for sample preparation, assembly of enzymatic reactions and electrophoresis. We have developed novel instrumentation designed to greatly increase the throughput of analyses of micro-satellites (short tandem repeat polymorphisms) in genomic DNA. This system is based upon Horizontal Ultrathin Gel Electrophoresis (HUGE) which employs denaturing polyacrylamide gels that are as thin as 50 microns.
The gel apparatus accommodates the parallel analysis of 144 lanes. An automated detection system scans the gels in real-time, collecting spectral information from four different fluorophores simultaneously. Typical separations yield excellent resolution of DNA fragments 300 nucleotides long in about 35 minutes. Reusable polyacrylamide gels that allow up to five electrophoresis runs on the same gel will also be discussed. In addition, we are developing an integrated robotics system to automate the assembly of the PCR mixture in microtiter plates, load the plates onto a thermal cycler, and automatically load the samples onto the gels.