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Figure 4 | Plant Methods

Figure 4

From: Implementation of two high through-put techniques in a novel application: detecting point mutations in large EMS mutated plant populations

Figure 4

Output data from HRM analysis (A and B). The upper panel shows the fluorescence change in dependence on the temperature. The lower panel shows the relative difference in melting curves compared to a reference sample. Decrease in fluorescence reflects the annealing state of the duplex species in the sample. Samples starting to melt at a lower temperature are likely to contain a SNP within the amplified fragment. (A) These graphs are presented the data from the serial dilution experiment. A PCR product (TG581 RFLP marker (SGN-M84)) has been amplified from S. lycopersicum (grey), S. pimpenellifolium (pink) or dilution of S. pimpenellifolium in S. lycopersicum with the following ratios: 1/1 (light blue); 1/3 (dark blue); 1/7 (green); 1/9 (red); 1/15 (blue); 1/31 (orange). (B) HRM screening output from screen performed on 4× pools from the M2 population for the PSY gene (PSY-1 fragment; table 1). The pink melting curve corresponds to a pool containing a C to T mutation within the amplified fragment, position and type of mutation was identified by Sanger sequencing.

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Plant Methods

ISSN: 1746-4811