INCREASED SPEED IN DNA SEQUENCING BY UTILIZING LARIS TO LOCALIZE MULTIPLE STABLE ISOTOPE LABELED FRAGMENTS

MAPPING AND SEQUENSING THE DNA IN THE HUMAN GENOME IS OF CRUCIAL IMPORTANCE TO FUTURE MEDICAL ADVANCES AND ECONOMIC COMPETITIVENESS. UTILIZATION OF DNA PROBES LABELED WITH DOZENS OF STABLE ISOTOPES INSTEAD OF A SINGLE RADIOISOTOPE COULD INCREASE THE RATE OF SEQUENCE DETERMINATION 100-FOLD OR MORE COMPARED TO CURRENT METHODS. THIS PROJECT INVOLVES A NEW APPROACH, LASER ATOMIZATION RESONANCE IONIZATION SPECTROSCOPY (LARIS), TO LOCALIZE AND QUANTIFY THE ISOTOPES IN DNA FRAGMENTS THAT HAVE BEEN SEPARATED BY POLYACRYLAMIDE GEL ELECTROPHORESIS. THE UNIQUE SELECTIVITY AND ULTIMATE SENSITIVITY MADE POSSIBLE BY THE LARIS TECHNIQUE ALLOWS DETECTION OF ISOTOPE TAGGED DNA FRAGMENTS IN THE 10(-18) MOLE OR LOWER RANGE WITHOUT ISOBARIC AND OTHER INTERFERENCES. THIS TECHNIQUE CAN BE APPLIED NOT ONLY TO GENOME SEQUENCING BUT ALSO TO HYBRIDIZATION METHODS USED IN GENETIC ENGINEERING. THIS PROJECT WILL USE THE SAME EXPERIMENTAL APPARATUS AS IN THE ONGOING SPUTTER INITIATED RESONANCE IONIZATION SPECTROSCOPY (SIRIS) DNA SEQUENCING MEASUREMENTS. THIS ALLOWS COMPARISON AND EVALUATION OF THESE TWO TECHNIQUES. DURING PHASE II, RECOMMENDATIONS WILLBE MADE FOR THE MOST FRUITFUL DIRECTIONS IN DNA SEQUENCING DEVELOPMENT. AN RIS SYSTEM DEDICATED TO DNA SEQUENCING WILLBE SPECIFIED.