Circular RNA Profiling by Illumina Sequencing via Template-Dependent Multiple Displacement Amplification
Divergent PCR for validation of NGS-tdMDA derived circRNA. Two products (>100bp and >200bp) were amplified with nb_circ7 primer upon divergent PCR (lane 4,5,6) with three N. benthamiana cDNA (a). With same primer, it did not give same size amplicon with genomic DNA (lane 1,2,3) as template (b). Two amplicons at ~100bp and 270 bp formed from rice cDNA (lane3) with osi_circ10 divergent primer whereas non-specific amplicon also formed when taking genomic DNA as template (lane 2) (c). Non-template PCR was taken as negative control (lane 2 in (a), lane 5 in (b), and lane 4 in (c)) and 100 bp amplicon formed from 5.8s as positive control (lane 1 in (a), lane 4 in (b), and lane 5 in (c)). Generuler 100 bp plus ladder in lane 3 (a), lane 6 (b), and Fermentas 100 bp ladder in lane 1 (c).