Puretaq ready to go pcr

Before the PCR-based analyses were performed, DNA was purified with GenElute Bacterial DNA kit (Sigma-Aldrich, St. Louise, MO, USA). For preparation of the PCR mixtures, PureTaq Ready-To-Go PCR (GE Healthcare; Buckinghamshire, UK) was used. For the sequencing procedure, DNA of amplified genes was sent to Eurofins MWG Operon (Ebersberg, Germany).

All isolates were serotyped. In addition, all serotype b isolates were leukotoxin promoter and AP-PCR genotyped. For the preparation of the PCR mixtures, PureTaq Ready-To-Go PCR (GE Healthcare; Little Chalfont, UK) was used. For serotyping and leukotoxin promoter typing, DNA was isolated by heating suspensions of the isolates in water for 8 min, while for the AP-PCR, the DNA was purified with GenElute Bacterial DNA kit (Sigma–Aldrich, St. Louis, MO).
The primers used and the PCR program for the serotyping and leukotoxin promoter typing are described elsewhere [29 (link)–31 (link)]. For the AP-PCR genotyping, the random sequence oligonucleotide OPB-3 (AGTCAGCCAC; Invitrogen, Carlsbad, CA; 0.4 µM) was used. The concentration of MgCl₂ in the PCR mixture was increased to 2.5 µM. The amplification was carried out as previously described [32 (link)].
The sequencing of the house-keeping gene, hbpA-2, was carried out as described elsewhere [18 (link)].