Gel electrophoresis, DNA purification and Sanger sequencing
All amplified samples were run on a 1.5% agarose gel and DNA products
of correct size were extracted using the QIAquick Gel Extraction Kit
(Qiagen, Hilden, Germany) following the manufacturer’s protocol.
Subsequently, extracted DNA was sent for Sanger sequencing using the
Microsynth Laboratory service (Microsynth, Göttingen, Germany). For thetp0488 gene product, and for the IGHGCH2 and the IGHG hinge
region amplicons, we utilized the respective forward primer for
sequencing. The tp0548 amplicons, however, were sequenced
bidirectionally using the internal sequencing primers published
elsewhere (Matějková et al., 2009).