MAMMALIAN DNA SEQUENCE MAPPING CENTER

BECAUSE OF THE RAPIDLY INCREASING USE OF IN SITU DNA SEQUENCE MAPPING TO MITOTIC CHROMOSOMES, A CENTER SHOULD BE ESTABLISHED TO DEVELOP THE MOST EFFICIENT TECHNIQUES AND TO AUTOMATE THE PROCEDURE. SUCH A CENTER WOULD MAKE AVAILABLE TO LABORATORIES INVOLVED IN THE CLONING OF DNA SEQUENCES THEABILITY TO EFFICIENTLY MAP TH SEQUENCES TO MAMMALIAN CHROMOSOMES. DNA SEQUENCING BY IN SITU HYBRIDIZATION WOULD THEREFORE BE MADE AVAILABLE TO LABORATORIES THAT DO NOT POSSESS THE TECHNICAL EXPERTISE IN HIGH-RESOLUTION CYTOGENETICS ESSENTIAL TO THIS MAPPING TECHNIQUE. CONDITIONS FOR IN SITU HYBRIDIZATION OF MOLECULAR PROBES TO MITOTIC CHROMOSOMES WOULD BE FURTHER DEVELOPED, FOCUSING ON IMPROVEMENTS IN PROBE LABELLING PROTOCOLS. COMPUTER- AUTOMATED PROCESSING FOR IN SITU HYBRIDIZATION AND AUTOMATEDDATA ACQUISITION AND ANALYSIS OF AUTORADIOGRAPHIC GRAIN DISTRIBUTIONS WOULD ALSO BE DEVELOPED. PHASE I ACTIVITY WILL BE CENTERED AROUND THE DEVELOPMENT OF COMPUTER PROGRAMSTO ANALYZE GRAIN DISTRIBUTIONS ON CHROMOSOMES AND THE STUDY OF THE AVAILABILITY OF COMPONENTS FOR MICRO-COMPUTER- CONTROLLED, AUTOMATED HYBRIDIZATION, AND AUTORADIOGRAPHIC PROCEDURES. THIS PHASE I ACTIVITY WOULD ALSO INCLUDE STUDIES OF THE OPTIMIZATION OF MOLECULAR PROBE LABELLING PROTOCOLS FOCUSING PARTICULARLY ON THE USE OF 35S-RNA LABELLING. FUTURE PHASE II ACTIVITIES WOULD INCLUDE CONSTRUCTION OF AN AUTOMATED IN SITU HYBRIDIZATION SYSTEM USING MICROCOMPUTER-CONTROLLED ROBOTICS TECHNOLOGY AND ENHANCEMENT OF THE COMPUTER ANALYSIS PROGRAMS TO PROVIDE ARCHIVING OF GENE SEQUENCE MAPPING DATA.