SCANNING SYSTEM TO MEASURE FLUORESCENCE FROM DNA GELS

THE OBJECTIVE IS TO DEVELOP A HIGH-SPEED, HIGH-RESOLUTION LASER SCANNING INSTRUMENT TO MEASURE BOTH THE MASS AND THE MOLECULAR WEIGHT OF DNA SEPARATED BY GEL ELECTROPHORESIS. THE INSTRUMENT DETECTS VISIBLE FLUORESCENCE FROM A DNA- SPECIFIC DYE EXCITED BY A VISIBLE CW LASER. EXPERIMENTS WITH A SLOW-SCANNING BREADBOARD INSTRUMENT HAVE DEMONSTRATED(1) A 100X IMPROVEMENT IN SENSITIVITY OVER PRESENT PHOTOGRAPHIC METHODS, AND (2) THE POTENTIAL FOR MEASURING SAMPLE FLUORESCENCE MORE ACCURATELY BY EMPLOYING IMPROVED TECHNIQUES TO CONTROL AND CORRECT FOR VARIATION IN BACKGROUND FLUORESCENCE. THE MASS OF DNA IN A BAND IS DIRECTLY PROPORTIONAL TO THE SUMMED FLUOREESCENCE SIGNAL. IN PHASE I, THE PRESENT BREADBOARD WILL BE MODIFIED TO SCAN100X FASTER WITH A HIGHER OUTPUT LASER IN ORDER TO SCAN A 20-CM-LONG GEL LANE IN UNDER 5 MINUTES. THE GREATER SPEED AND DATA-HANDLING CAPACITY WILL PERMIT PERFORMING THE EXTENSIVE TESTING REQUIRED TO MORE FULLY ESTABLISH INSTRUMENTAL AND ANALYTICAL PERFORMANCE, AND STUDYING FURTHER SAMPLES IN SEVERAL TYPES OF GELS. AN ENGINEERING PROTOTYPE WILL BE BUILT AND EXTENSIVELY TESTED IN PHASE II THAT WILL USE A HIGHER POWER LASER, IMPROVED DATA-HANDLING TECHNIQUES, AND OTHER IMPROVEMENTS TOFURTHER INCREASE MEASUREMENT SPEED AND AUTOMATION. AWARDEE'S STATEMENT OF THE POTENTIAL COMMERCIAL APPLICATIONSOF THE RESEARCH: SIGNIFICANTLY IMPROVED ANALYTICAL PERFORMANCE, THE PROSPECT OF INCREASED AUTOMATION, AND LOW TO MODERATE PRICE ARE HIGHLY ATTRACTIVE FEATURES OF THE INSTRUMENT, BOTH FOR NUCLEIC ACID AND OTHER APPLICATIONS IN GEL ELECTROPHORESIS. A POTENTIALLY LARGE COMMERCIAL MARKET EXISTS FOR AN IMPROVED LASER SCANNING INSTRUMENT.