Denovo sequencing of Ocimum sanctum Genome sequencing
2 microgram of genomic DNA prepared in nuclease free water (Ambion) was prepared. To prepare the DNA library DNA was sonicated (Covaris), to fragment length ranging between 150 to 600 bp. The resulting fragmented DNA was cleaned up using Agencourt Ampure XP SPRI beads (Beckman Coulter). The size distribution was checked by running an aliquot of the sample on Agilent High Sensitivity Bioanalyzer Chip.  Subsequently libraries for whole genome sequencing were constructed according to the TruSeq DNA library protocol outlined in “TruSeq DNA Sample preparation guide” (Part # 15005180; Rev. A; Nov 2010). DNA was subjected to a series of enzymatic reactions that repair frayed ends, phosphorylate the fragments, and add a single nucleotide A overhang and ligate adaptors (Illumina’s TruSeq DNA sample preparation kit). Sample cleanup was done using Ampure SPRI beads. After ligation, ~500 – 600 bp for long insert, and 300-400bp for short insert were size selected on a 2% agarose gel and cleaned using MinElute column (QIAGEN). PCR amplification was done and cleaned up using Ampure SPRI beads.
CIMAP_ePCR1_Long