The genomic DNA of CP118 was manually extracted from the strain cultured on modified GAM agar (Nissui Pharmaceutical Co.) supplemented with 5% sheep blood at 37°C for 24 hr under anaerobic
conditions according to the methods described by Okamoto et al. [37 (link)]. In brief, CP118 was suspended in TE (10 mM Tris–HCl [pH 8.0] and
1 mM EDTA [pH 8.0]), treated with lysozyme and mutanoysin, and lysed by 10% sodium dodecyl sulfate. The lysate was then treated with phenol, phenol-chloroform-isoamyl alcohol (25:24:1)
(PCI), and chloroform once, three times, and twice, respectively. Nucleic acid was precipitated and washed by ethanol and resuspended in sterile H2O. After the RNase treatment and
additional extraction with PCI and chloroform, bacterial DNA was precipitated again, rinsed with ethanol, and dissolved in 10 mM Tris-HCl (pH 8.5).
conditions according to the methods described by Okamoto et al. [37 (link)]. In brief, CP118 was suspended in TE (10 mM Tris–HCl [pH 8.0] and
1 mM EDTA [pH 8.0]), treated with lysozyme and mutanoysin, and lysed by 10% sodium dodecyl sulfate. The lysate was then treated with phenol, phenol-chloroform-isoamyl alcohol (25:24:1)
(PCI), and chloroform once, three times, and twice, respectively. Nucleic acid was precipitated and washed by ethanol and resuspended in sterile H2O. After the RNase treatment and
additional extraction with PCI and chloroform, bacterial DNA was precipitated again, rinsed with ethanol, and dissolved in 10 mM Tris-HCl (pH 8.5).
