MULTIPLEX GENOMIC WALKING: INTEGRATION OF THE WET LAB AND COMPUTER LAB INTO A SINGLE PROTOTYPING ENVIRONMENT

We are presently sequencing the entire genome of Mycoplasma capricolum, one of the smallest of free living organisms by a Multiplex Genomic Walking strategy. This technique involves the repetitive hybridization of sequencing membranes with oligonucleotide probes to acquire sequence data in discrete steps along the genome. The technique allows one to walk a genome in a directed manner eliminating the problems associated with random shotgun assembly. Furthermore, the repetitive stripping and hybridization process is relatively simple to reproduce and has the potential to be easily automated. The Genetic Data Environment (GDE), an X Windows based Graphic User Interface has allowed the seamless integration of a core multiple sequence editor with pre-existing external sequence analysis programs and internally developed programs into a single prototypic environment. This system has facilitated linkage of the Harvard Genome Lab's internal database and automated data control systems into one Graphic User Interface which can handle the archiving and analysis of both random fluorescent sequencing data and genomic walking data from the Mycoplasma project. Finally, it has facilitated the integration of the Genomic sequence data into a PROLOG database environment for the comparative analysis of Mycoplasma capricolum and other organisms.