De novo Genome sequencing of stalk infecting fungal pathogen Colletotrichum falcatum
Libraries for whole genome sequencing was 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. Sample cleanup was done using Ampure SPRI beads. After ligation, ~300 – 500 bp fragment was size selected on a 2% agarose gel and cleaned using MinElute column (QIAGEN). PCR (10 cycles) amplification of adaptor ligated fragments was done and cleaned up using Ampure SPRI beads. The prepared libraries were quantified using Nanodrop and validated for quality by running an aliquot on High Sensitivity Bioanalyzer Chip (Agilent).