SBIR-STTR Award

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 the ability to efficiently map the 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 highresolution 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 automated data acquisition and analysis of autoradiographic grain distributions would also be developed.Phase I activity will be centered around the development of computer programs to analyze grain distributions on chromosomes and the study of the availability of components for microcomputer-controlled, automated hybridization, and autoradiographic procedures. This Phase I activity would also include studies of the optimization of molecular probe labelling pyotocols focusing particularly on the use of 35SRNA 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.National Institute on Aging (NIA)

Because of the rapidly increasing utilization of in situ DNA sequence mapping to mitotic chromosomes, it is proposed to automate the procedure and to employ efficient microcomputer-assisted mapping analysis techniques. This project would make available to laboratories involved in the analysis of mammalian genomes the capability of efficiently mapping these sequences to mammalian chromosomes. DNA sequence mapping by in situ hybridization would, therefore, be available to laboratories that do not possess technical expertise in high-resolution cytogenetics essential to this mapping technique. Computer-automated DNA probe processing for in situ hybridization and automated data acquisition and analysis of gene probe distributions are the major technical objectives of this project. Phase I activity centered around the development of computer programs for the analysis of probe distributions on chromosomes and prototype instrument development for microcomputer-controlled automated hybridization procedures. Phase II activity will include construction of an automated in situ hybridization system using microcomputer-controlled technology, as well as enhancement of the mapping analysis system to provide efficient DNA sequence mapping data collection and management.

Anticipated Results:
In situ hybridization has been increasingly used as a tool for genome analysis. Access to a commercial venture that would offer this capability would be valuable to molecular biologists who lack the experience, the time, or the resources to do such studies. Alternatively, automated mapping instrumentation could be made available to laboratories with cytogenetics expertise. The instrument would also be adaptable to other microscope slide-based analytical techniques.National Institute on Aging