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