PLASTIC FILM-SUPPORTED DNA SEQUENCING GELS

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 SUPPORTINGMATERIAL. 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 FORDNA 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 PLANDESCRIBES 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 DESIGNAND CONSTRUCTION OF A SYSTEM TO MANUFACTURE PRECAST GELS. ULTIMATELY, PLASTIC FILM-SUPPORTED GELS WILL BE SOLD FOR USEIN BETAGEN CORPORATION'S AUTOMATED DNA-SEQUENCING INSTRUMENTS AND MOST MANUFACTURERS' ELECTROPHORESIS SYSTEMS.