IMPROVED REPLACEMENT TECHNOLOGY FOR RFLP ANALYSIS

RFLP ANALYSIS HAS BEEN SHOWN TO HAVE TREMENDOUS IMPLICATIONSFOR BOTH THE DIAGNOSIS OF GENETIC TENDENCIES FOR HUMAN DISEASE AND FOR APPLICATIONS TO PLANT AND ANIMAL IMPROVEMENT; UNFORTUNATELY, THE DEMANDS FOR ITS USE HAVE OUTSTRIPPED THE CAPACITY OF WHAT IS PRIMARILY A RESEARCH TECHNOLOGY. A REPLACEMENT TECHNOLOGY IS DESIRED THAT IS FASTER, CHEAPER, AND WITH A HIGHER THROUGHPUT BUT, AT THE SAME TIME, YIELDS COMPARABLE INFORMATION. CERES/NPI HAS DEVELOPED A PCR-BASED APPROACH THAT APPEARS TOSATISFY MANY OF THESE CRITERIA. THIS PROCESS DELIVERS INFORMATION AS TO THE DNA SEQUENCE VARIABILTIY BETWEEN INDIVIDUALS AT SPECIFIC LOCI, WHICH CAN THEN BE USED ANALOGOUSLY TO RFLP RESULTS. REDUCTION TO PRACTICE REQUIRESTHAT A BETTER UNDERSTANDING OF AMPLIFICAITON KINETICS BE DEVELOPED AND EXACT CONDITIONS FOR SEQUENCE DISCRIMINATION BE FURTHER REFINED; IMPLEMENTATION OF A CAPTURE-SIGNAL STRATEGY IS ESSENTIAL TO REMOVE THE NECESSITY FOR RADIOACTIVE LABELS AND GEL ANALYSIS. THE RESULTING PROCESS SHOULD REQUIRE LESS THAN A DAY FOR COMPLETION, BE CHEAPER BY 10-FOLD OVER CONVENTIONAL ANALYSIS, AND FACILITATE THE ANALYSIS OF SEVERAL LOCI FOR MANY SAMPLES, ALL IN A LABORATORY WITH PERSONNEL MUCH LESS SOPHISTICATED THAN NOW REQUIRED. AWARDEE'S STATEMENT OF THE POTENTIAL COMMERCIAL APPLICATIONSFOR THE RESEARCH: IF THIS PROCESS CAN BE REDUCED TO PRACTICE BY TAKING ADVANTAGE OF MANY OF THE ADVANCES IN OTHER AREAS OF DIAGNOSTIC TECHNOLOGIES, IT SHOULD REALIZE THE SAME GOAL AS IS NOW ACCOMPLISHED BY THE MORE CUMBERSOME RFLP-BASED METHODOLOGY BUT SATISFY THOSE DEMANDS EXPECTED OFAN APPLIED TECHNOLOGY WITH MANY DIFFERENT UTILIZATIONS IN HUMAN DIAGNOSTICS, PLANT AND ANIMAL BREEDING, AS WELL AS MANY DIFFERENT RESEARCH APPLICATIONS.