THE TECHNIQUES OF DNA SEQUENCING ARE OF CENTRAL IMPORTANCE IN MOLECULAR BIOLOGY AND BIOTECHNOLOGY. DETERMINATION OF THE COMPLEX NUCLEOTIDE SEQUENCES OF LARGE MOLECULES BY THE PRESENT MANUAL METHODS IS, HOWEVER, A TEDIOUS AND TIME CONSUMING PROCESS AS ARE THE ITERATIVE MANUAL DETERMINATIONS OF RELATIVELY SHORT SEQUENCES. THE REPETITIVE NATURE OF THE TASK MAKES IT AN IDEAL SUBJECT FOR AUTOMATION.
THE AUTOMATION THE PROCESS AIMED AT IN THE PRESENT PROPOSAL SHOULD SPEED UP THE DETERMINATION OF NUCLEOTIDE SEQUENCES FOR THE BENEFIT OF BOTH ACADEMIC AND INDUSTRIAL RESEARCH.
An imaging system has been produced, based on gas wire counter technology, which is suitable for imaging the beta distributions encountered in blotted autoradiographs. Data is captured and displayed on an IBM PC/AT with a dedicated graphics system. The chief benefits of the apparatus are the large decrease in exposure times (approximately 1/20) required for data measurements relative to X-ray film and the availability of quantitative data.
Deoxyribonucleic acid (DNA) sequencing is an obvious candidate for automation allowing more rapid access to the genetic data encoded in very large DNA molecules, such as the human genome.
Research was carried out into the production of fast and accurate instrument modules to automate aspects of sequence analysis. A novel machine was designed and built which incorporated a vision controlled robot to identify and select plaques and colonies on Petri dishes for automatic sampling and transfer of candidate material for further growth in individual containers. Electrophoresis and direct blotting were automated and a gas counter was built for directly imaging radiolabelled DNA sequences and mapping gels for genetic data abstraction.
The first robot system (APSCIR) to image mixed randomly disposed arrays of biological specimens and to pick a particular set of these for distribution into an ordered arrangement was produced. Imaging hardware and software were developed jointly with a patented robot picking head to automate this process, which is recognized as a serious bottleneck in many DNA analysis procedures. The direct blotting electrophoresis system was developed into a key technology for the second generation of automatic mappers and sequencers and the gas counter capabilities were extended into the area of sequence and genetic analysis.
TRANSFER OF ELECTROPHORESIS GEL DATA TO A NEW MULTIWIRE PROPORTIONAL COUNTER (MWPC).
THE MWPC WILL BE MODIFIED TO ACCEPT THE BLOTTING MATRIX PRODUCED BY THE COLLABORATORS FROM THE UNIVERSITY OF CONSTANCE.
SOFTWARE DEVELOPMENT FOR MWPC IMAGE ANALYSIS ON AN IBM PC/AT AND ABSTRACTION OF SEQUENCE DATA FROM DNA BAND PATTERNS.
Funding SchemeCSC - Cost-sharing contracts
M60 1QD Manchester