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. 2020 Sep 30;13:496. doi: 10.1186/s13071-020-04371-0

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© The Author(s) 2020

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — Lower limit detection and optimum primer concentration of G. truncatula LAMP assay. This was determined by undertaking (a) LAMP assay and (b) confirmation of results of the LAMP assay products using agarose gel (2%) electrophoresis. Lane M: 100 bp DNA Ladder (Thermo Fisher Scientific). Lane + (positive control of 34.9 ng/μl G. tuncatula genomic DNA extract) and Lane− (negative control) were both undertaken at primer concentration 3 pmol of F3 and B3 primer, 25 pmol of FIP and BIP primer and 12 pmol loop B and loop F primer. Lane 1 (10⁴ dilution of G. tuncatula genomic DNA extract), Lane 2 (10⁵ dilution of G. tuncatula genomic DNA extract) and Lane 3 (10⁶ dilution of G. tuncatula genomic DNA extract) were undertaken at primer concentration 2.5 pmol of F3 and B3 primer, 22 pmol of FIP and BIP primer and 10 pmol of loop B and loop F primer. Lane 4 (10⁴ dilution of G. tuncatula genomic DNA extract), Lane 5 (10⁵ dilution of G. tuncatula genomic DNA extract) and Lane 6 (10⁶ dilution of G. tuncatula genomic DNA extract) were undertaken at primer concentration 3 pmol of F3 and B3 primer, 25 pmol of FIP and BIP primer and 12 pmol of loop B and loop F primer. Lane 7 (10⁴ dilution of G. tuncatula genomic DNA extract), Lane 8 (10⁵ dilution of G. tuncatula genomic DNA extract) and Lane 9 (10⁶ dilution of G. tuncatula genomic DNA extract) were undertaken at primer concentration 3.5 pmol of F3 and B3 primer, 28 pmol of FIP and BIP primer and 14 pmol of loop B and loop F primer