SEPARATION MEDIA FOR DNA SEQUENCING

HIGH-SPEED, HIGH-THROUGHPUT DNA SEQUENCING METHODS RELY ON ELECTROPHORETIC SEPARATION IN VERY NARROW GEOMETRIES, I.E., MICRO-CAPILLARIES AND ULTRA-THIN SLABS. AS CHANNEL DIMENSIONS DECREASE, THE SEPARATION MEDIUM BECOMES A CRITICAL, LIMITING FACTOR IN THE SPEED, ACCURACY, AND REPRODUTIBILITY OF DNA FRACTIONATIONS. CONVENTIONAL, IN-SITU CASTING OF GELS IS NOT WELL SUITED FOR CONFINED GEOMETRIES, AND THE CURRENT APPROACH OF USING UN-CROSSLINKED POLYMER SOLUTIONS HAS SEVERAL PERFORMANCE DEFICIENCIES, INCLUDING RESOLUTION AND REPRODUCIBILITY LIMITATIONS. THIS PROJECT INVOLVES A COMPLETELY NOVEL SEPARATION MEDIA CONCEPT WHICH COMBINES THE DESIRABLE PERFORMANCE ATTRIBUTES OF CROSSLINKED NETWORKS, THE LOADING/UNLOADING FEATURE OF DILUTE POLYMER SOLUTIONS, AND THE USER CONVENIENCE OF PRE-CAST GELS. THE NEW APPROACH WILL UTILIZE DISCRETE, 'SMART' GEL PARTICLES WHICH FORM A FREE-FLOWING PSEUDO-NETWORK UNDER APPROPRIATE CONDITIONS, AND CAN BE READILY INJECTED INTO, AND FLUSHED FROM, NARROW CAPILLARIES. THE FEASIBILITY OF THIS DNA SEPARATION IN A CAPILLARY FORMAT.