The whole genome analysis of endophytic bacteria of maize seed varieties elucidating immune response to Fusarium graminearum
The library was prepared using KAPA HyperPlus Kits (Roche) following the manufacturer's user guide. The initial concentration of DNA was evaluated using the Qubit dsDNA HS Assay Kit (Life Technologies). 25 ng DNA was used to prepare the library. Protocol starts with enzymatic fragmentation to produce dsDNA fragments followed by end repair and A-tailing to produce end-repaired, 5'-phosphorylated, 3'-dA-tailed dsDNA fragments. In adapter ligation step, dsDNA adapters are ligated to 3'-dA-tailed molecules. Final step is library amplification, which employs high fidelity, low-bias PCR to amplify library fragments carrying appropriate adapter sequences on both ends. The library was diluted (to 10.0 pM) and sequenced paired end for 500 cycles using the HiSeq system (Illumina).