Genomic DNA Isolation and SSR Genotyping

Genomic DNA was isolated from leaf tissue of 16 randomly selected individuals from each population. A set of 720 DNA samples (45 populations × 16 individuals) were isolated along with the control sample Tift 23D₂B₁ using NucleoSpin® 96 Plant II Kit (Macherey-Nagel, Germany). Electrophoresis (0.8% agarose gel) was performed to test the quality of the DNA and quantified based on lambda DNA (MBI Fermentas, USA). The final working DNA samples were normalized uniformly at a concentration of 10 ng/µl.
Twenty-nine SSR markers (Supplementary Table 2), identified as highly polymorphic and distributed over all the seven linkage groups based on earlier studies^{66 (link),67 (link)} were used. The detailed methodology followed for DNA extraction, SSR genotyping protocol and allele calling using Genemapper 4.0 (Applied Biosysterms) is explained in Patil et al.⁴⁶ .