Arka Vikas (sel.22)
DNA samples were subjected to quality control and considered for library preparation using the standard protocols (Peterson et al., 2012). 1 g genomic DNA of each of the twelve tomato lines was double digested with SphI and MlucI restriction enzymes and incubated at 37 C for 16-20 hours. Cleaning the unwanted restricted fragments using AMpure XP beads (Beckman Coulter Genomics). Ligation of P1 (Barcoded) and P2 adapters was carried out using T4 DNA ligase. Cleaning the unwanted ligated fragments using AMpure XP beads(Beckman Coulter Genomics). Ligated product was fractionated in 2% SybrSafe agarose gel electrophoresis to check the product size in between 250 - 350 bp.  To increase the concentration of sequencing libraries, PCR amplification (8-12 cycles) was performed  by adding Index 1 and Index 2 (8 nt long) for multiplexing using PhusionTMTaq DNA polymerase kit.  Products of PCR amplification was analyzed in Agilent Bioanalyzer to quantify molarity and fragment size distribution. For each sample, individual libraries were prepared and included in one lane. The sequencing was done based on V4 Chemistry on HiSeq2500 platform