A MICROCOMPUTER-BASED DNA GEL ELECTROPHORESIS IMAGE ANALYSIS

THIS PROJECT IS TO DESIGN AND CONSTRUCT A MICROCOMPUTER-BASED INSTRUMENT FOR QUANTITATING SIZE DISTRIBUTIONS OF NUCLEIC ACIDS (DNA AND RNA) SEPARATED ACCORDING TO SIZE BY GEL ELECTROPHORESIS. THE STARTING POINT FOR ANALYSIS OF FLUORESCENCE-STAINED DNA IN GELS WILL BE THE IMAGE DIGITIZATION OF THE ENTIRE GEL ELECTROPHORETIC PATTERN (OR SPECIFIED PORTIONS OF IT) SO THAT THE RELATIVE INTENSITIES OF FLUORESCENCE WILL BE CONVERTED DIRECTLY TO IMAGE DATA IN COMPUTER MEMORY. ONCE THE GEL FLUORESCENCE IMAGE HAS BEEN DIGITIZED, THE IMAGE PROCESSING SYSTEM WILL PERFORM QUANTITATIVE MEASUREMENTS OF THE AMOUNT OF DNA OF VARIOUS SIZES USING SEMIAUTOMATED COMPARISON TO DNA SIZE STANDARDS. ADVANTAGES OF SUCH A SYSTEM INCLUDE FLEXIBILITY, PRECISION, AND RELIABILITY IN THE ANALYSIS; ACCEPTANCE OF VARYING FORMATS FOR GEL ELECTROPHORESIS; AND BECAUSE THE SYSTEM IS MICROCOMPUTER-BASED AND PROBABLY CAN BE ASSEMBLED FROM STOCK COMPONENTS, RELATIVELY LOW COST. PHASE I WILL FOCUS ON INVESTIGATIONS OF HARDWARE COMPONENT REQUIREMENTS, ASSEMBLY OF A PRELIMINARY HARDWARE DESIGN TO DEMONSTRATE FEASIBILITY OF THE PROPOSED CONFIGURATION FOR A WORKABLE GEL-DIGITIZATION ANALYSIS INSTRUMENT, AS WELL AS INITIAL SOFTWARE DEVELOPMENT TO DEMONSTRATE THE POTENTIAL ANALYTICAL UTILITY OF SUCH A SYSTEM. PHASE II WILL THEN INVOLVE CONSTRUCTION, THOROUGH TESTING AND VALIDATION OF THEOVERALL HARDWARE SYSTEM DESIGN, AND COMPLETE GENERATION OF ASOFTWARE SYSTEM FOR THE ACCURATE AND VERSATILE ANALYSIS OF DNA SIZE DISTRIBUTIONS ON GELS. SYSTEM DESIGN WILL PROVIDE THE BROADEST AND MOST VERSATILE SET OF DNA ANALYTIC FUNCTIONS POSSIBLE, TO ACCOMMODATE NEW APPLICATIONS IN MOLECULAR GENETICS, GENE MAPPING AND SEQUENCING, AND GENETIC TOXICOLOGY.