Description and application of a rapid method for genomic DNA direct sequencing

We describe a rapid method for determining nucleotide sequences directly fr om total genomic DNA. This technique was used to determine genomic DNA sequ ences in various prokaryotic and eukaryotic microorganisms with a G+C conte nt between 40 and 50%, e.g. Escherichia coli. Yibrio cholerae, Bacillus sub tilis and Saccharomyces cerevisiae. Furthermore, the method was applied to accurately sequence up to 300 DNA base pairs in Photorhabdus luminescens. w hose genome sequencing is currently under way. Taken together. these result s provide evidence that our technique can be widely used to easily and effi ciently determine genomic DNA sequences. (C) 2001 Federation of European;Mi crobiological Societies. Published by Elsevier Science B.V. All rights rese rved.