(A) CircRNA was synthesized by in vitro transcription from a permuted intron-exon template. RNase R treatment of the transcription products enriched for circRNA. Green arrows represent the convergent qRT-PCR primers to detect both linear and circRNA; brown arrows represent the divergent qRT-PCR primers to detect circRNA.
(B) Linear and circRNA were treated with RNase R and analyzed by Bioanalyzer.
(C) Sequencing of circRNA product reveals precise and uniform splice junction.