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. 2023 Sep 11;8:341. doi: 10.1038/s41392-023-01561-x

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — The hypothesis of purification results for group I intron permuted intron‒exon (PIE) system by E-Gel EX Agarose Gels. Splicing reaction by group I intron PIE system can produce several impurities. RNase R can digest a portion of linear RNA and nicked circRNA. Subsequently, HPLC can eliminate most of the fragments except for impurities of the same size. In addition, RNase H is the best option to detect the purity of circRNA products. The pure circRNA products exhibit only one bond and impure circRNA products exhibit two bonds