Fig. 2

Verification of linearity of the S. endobioticum mtDNA. a Design of the verification experiments using a graphical representation of the 5′ and 3’ TIRs (orange) and forward and reverse primer sites (green and light green). Three PCR reactions, performed after TdT tailing, are displayed: ❶ M13F_polyG/mtDNA_00787-fw to verify linear conformation of the mtDNA. This reaction takes place at both telomeric ends since the TIRs are inverse orientated. ❷ mtDNA_0007-fw/mtDNA_03691-rv to verify presence of the TIR with its specific flanking sequence on the 5′ end of the mtDNA. ❸ mtDNA_0007-fw/mtDNA_69209-fw to verify presence of the TIR with its specific flanking sequence on the 3′ end of the mtDNA. b Gel images corresponding with reactions described under A. The 1 kb-plus size marker (M) was used for amplicon size estimation. c Sanger sequence data mapped to the S. endobioticum mtDNA corresponding with the reactions described under A. The mtDNA is used as reference, and bases similar to the reference are shown in grey, and differences to the reference are highlighted (black). Phred scores for the individual peaks are shown as blue bars, and low quality data (Phred < 30) is annotated in pink. For reaction 1, the first 800 bases of the 5′ end TIR are shown, whereas for reactions 2 and 3 the full TIR including the specific flanking sequences are shown (~ 4 kb)