The purpose of this study was to detect the circRNA content in various organs of Nymphaea minuta
Materials and methods
After comparison with the reference genome, CircRNA was identified with unmatched Reads, which could not be directly compared with the reference genome because they came from distant exon regions and had large fragment gaps during direct comparison. As shown in the figure below, Reads from the linear RNA splicing site are aligned to the genome in the same position order as those in linear RNA, while Reads from the CircRNA junction point are aligned to the reference genome in the opposite position order. In order to distinguish Reads from linear RNA clipped sites and CircRNA junction sites, a portion of the sequence (called 5' Anchor and 3' Anchor, respectively) from the 5' end and 3' end of Reads that are inferior to the reference genome needs to be re-matched with the reference genome. If the two sequences are aligned in opposite locations, the Reads may come from a CircRNA. The Anchor sequence was extended all the way. If the sequence could match the reference genome completely up to the junction point, and the shear pattern at the junction point was consistent with the shear pattern of AG-GT, it was identified as a CircRNA.
