The overall contiguity of the Saccharomyces cerevisiae chromosome VI sequence assembly was assessed by systematic long-range PCR, PCR product size determination and sequencing. Using S. cerevisiae strain FY1679 total genomic DNA as template, 41 overlapping PCR products were generated, covering the complete 270 kb chromosome VI sequence. Specificity of the PCR products was confirmed by direct end-sequencing. No fragment size discrepancies with the published sequence were observed, confirming the overall sequence assembly. Gel reads of the PCR-fragment ends compile to a total of resequenced DNA representing 16% of the entire chromosome VI and reveal three single nucleotide differences. One of these is an extra G in the protein-coding region of YFL007w. Due to this additional nucleotide, the coding sequences of YFL007w, and YFL006w become part of a 6432 bp ORF. The same sequence also resulted from analysis of strain BY4743. Homologous proteins of unknown function found in Candida albicans, Arabidopsis thaliana, Caenorhabditis elegans and man, as well as comparative data from published transcript profiles of YFL006w and YFL007w, give additional evidence for the existence of a single gene at this locus in yeast. Copyright (C) 2002 John Wiley Sons, Ltd.