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. 2017 Sep 27;8:1678. doi: 10.3389/fpls.2017.01678

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Copyright © 2017 Chen, Cui, Wang, Zhu, Lu and Jin.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Various experimental strategies validated the circRNAs in A. thaliana. (A) A representative example of A. thaliana circRNAs (Ath_circ_FC4757) showing the validation strategy. Divergent and convergent primers were designed to detect circRNAs, while divergent primers were designed using the “out-facing” strategy. Sanger sequencing further confirmed head-to-tail back-splicing. (B) Divergent primers successfully amplified Ath_circ_FC4757, Ath_circ_FC4468, and Ath_circ_FC2295 in cDNA, but failed to do so in genomic DNA. Convergent primers worked on both cDNA and genomic DNA. actin 2: linear control.