Figure 4

High resolution melting peaks and amplicon sequences from Category 1 candidate SNPs between M. sativa subs. sativa var 'Chilean' (red) and M. sativa subsp. falcata var 'Wisfal' (gray). SNPs predicted by 454 sequencing are shown in blue and additional SNPs identified through amplicon sequencing are shown in black. A) Primers SNP MSSNPV002 confirms the presence of a single SNP at position 52 (Chilean = G, Wisfal = A) in a 106 bp amplicon. B) Primers SNP MSSNPV008 confirms the presence of SNP at position 68 (Chilean = G, Wisfal = C), and additional SNPs at position 48 (Chilean = G, Wisfal = C/G) and position 99 (Chilean = A/G, Wisfal = A) in a 138 bp amplicon. C) Validated candidate SNP MSSNPV009 at position 68 (Chilean = C, Wisfal = G), and additional SNPs at position 48 (Chilean = A/C, Wisfal = A), position 87 (Chilean = A/G, Wisfal = A/G), position 90 (Chilean = C/T, Wisfal = C/T), position 99 (Chilean = A/G, Wisfal = A/G), position 100 (Chilean = G, Wisfal = A/G), and position 155 (Chilean = A/G, Wisfal = A/G) in a 220 bp amplicon. D) Non-validated candidate SNP MSSNPV001 (Chilean = G, Wisfal = G) in a 136 bp amplicon.