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. 2022 Jul 29;12:13089. doi: 10.1038/s41598-022-17382-x

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© The Author(s) 2022

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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2% Agarose-TAE gel displaying the specificity of multiplex primers in the case of duplex formation in freshly extracted colony-reared larval DNA with species-specific amplicon size indicated. Lane 4 demonstrates the expected C. cosyra amplicon at 183 bp with non-specific amplification at 128 bp leading to the formation of a duplex; the larger 183 bp amplicon should be used for identification. Lane 5 is a single 128 bp amplicon indicative of C. quilicii. Lanes 1 & 2: C. capitata, Lanes 3 & 4: C. cosyra, Lanes 5 & 6: C. quilicii, Lanes 7 & 8: C. rosa, Lanes 9 & 10: B. dorsalis, Lane 11: No template control, Lane L: GeneRuler 100 bp DNA ladder (Thermo Scientific). The original gel is presented in Supplementary Fig. S2a.