Fig. 3

Validation of novel circRNAs corresponding to the 5′-part of the homologous transposase genes SSO_RS05855 and SSO_ RS06560. a and b Schematic representations of divergent primers for detection of circular RNA or convergent primers for detection of linear and circular RNA. Grey lines: RNA. Black arrows: oligonucleotides (primers) for RT-PCR. Short black line: circularization junctions. c and d Ethidium bromide-stained 10% polyacrylamide gels showing products from RT-PCR analyses of SSO_RS05855 and SSO_RS06560 (indicated). The use of divergent or convergent primers is given below the panels. RNA (− R): total RNA was used as template for cDNA synthesis by reverse transcriptase and then PCR was performed (RT-PCR analysis). RNA (+ R): total RNA treated with RNase R (which degrades linear but not circular RNA) prior to RT-PCR. H₂O and DNA: positive and negative PCR control reactions, to which water or total DNA was added instead of RNA. M: length marker (size is given in bp). Uncropped images of c) and d) are shown in Fig. S1 (Additional file 1)