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. 2020 Sep 15;10:15144. doi: 10.1038/s41598-020-71614-6

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© The Author(s) 2020

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Overview of the protocol developed in this study. DNA was sheared into 6 kb fragments, end-repaired and circularized. An inverse long-range PCR, using p35S primers orientated in the reverse direction, specifically amplified circularized DNA molecules that contained the p35S promoter. PCR products containing the transgene or a portion of the transgene were sequenced using the Oxford Nanopore sequencer. The bioinformatic pipeline allowed to recover the sequence of the transgene and its flanking regions.