Figure 1

PCR and Southern hybridization to confirm deletion genotypes. PCR and Southern hybridizations were performed on genomic DNA isolated from the indicated strains. (A) For uvrA, PCR will amplify a 3022 bp fragment if the wild-type allele is present and a 195 bp fragment if the deletion allele is present. For uvrC, presence of the wild-type allele is indicated by amplification of an 1857 bp fragment, the deletion allele by amplification of a 1017 bp fragment. Lanes 1 and 10 contain Hyperladder I for reference (BioLine). PCR primers are listed in Table 3. Similar results were obtained using primers for uvrB (data not shown). (B) Genomic DNA was digested with PstI and, following electrophoresis and transfer to a charged membrane, was hybridized with chemifluorescently labelled PCR fragments to a region upstream of the uvrA gene (top) or uvrB gene (bottom). Similar results were obtained for blots hybridized with probes to the uvrC genomic region (data not shown).