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PCR amplification was carried out with PCR primers (20 pmol/L each per 50 [micro]L reaction) (Integrated DNA Technologies, Coralville, IA, USA) (Table) and 1 [micro]L genomic DNA (extracted from overnight Luria-Bertani broth cultures according to PureGene DNA isolation kit instructions [Gentra Systems, Minneapolis, MN, USA] and dissolved in 50 [micro]L 10 mmol/L Tris, pH 8. Portions of these frozen stocks were transferred to vancomycin-supplemented (20 mg/L) Luria-Bertani broth. The bacteria were grown while being shaken overnight at 37[degrees]C in Luria-Bertani broth (LB). |
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