The objective of the research plan is to develop a process of preparing thin polyacrylamide gels for DNA sequencing, substituting plastic film for glass plates as the supporting material. A process to make plastic film-supported gels with satisfactory electrophoretic properties will allow preparation of ready-to-use gels and form the basis of a useful manufacturing process. Commercial availability of preformed gels will eliminate manual preparation of gels for DNA sequencing, a time-consuming and tedious step common to all DNA-sequencing procedures.Initial studies show that gels formed on a variety of plastics do not provide the resolution needed for DNA sequencing. Evidence suggests incomplete acrylamide polymerization at the gel/ film boundary. This research plan describes several experimental approaches aimed at ensuring complete gel polymerization on polyester films. The protocol used for preparation of high-resolution, plastic film-supported gels developed in Phase I of this project will be the foundation of a Phase II proposal for the design and construction of a system to manufacture precast gels. Ultimately, plastic film-supported gels will be sold for use in Betagen Corporation's automated DNA-sequencing instruments and most manufacturers' electrophoresis systems.National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
