Jetpei carrier

For polymerase chain reaction (PCR)-optimization and in vivo transfection by JetPei-carrier (Polyplus transfection, Illkirch, France) [13 (link)], we first needed to liberate whole PCV2 genotype group members from each hybrid plasmid, pCR2.1-TOPO (pCR2.1-TOPO, Invitrogen, Basel, Switzerland) by EcoR I (New England Biolabs, Bioconcept, Allschwil, Switzerland) digestion and purification over a size exclusion column (MinElute PCR Purification column, Qiagen; Hombrechtikon, Switzerland). As intramolecular reactions are faster than intermolecular reactions, we directly ligated (T4-DNA Ligase; New England Biolabs, Allschwil) the DNA mix from the plasmid backbone and whole PCV2 group member digestion/purification. This ligation was again purified over a column (MinElute PCR Purification column, Qiagen; Hombrechtikon, Switzerland). The resulting whole PCV2 group member DNA ligation mix was used as a template for optimizing the PCR reaction conditions and in vivo transfection experiments, as described in Klausmann et al. [13 (link)].