title: DNA-seq of Bacillus tequilensis
design: Library preparation was performed  using  NEXTFlex DNA library protocol outlined in NEXTFlex DNA sample preparation guide (Cat # 5140-02). In brief, genomic DNA was sheared to generate fragments of approximately 300-500bpin a CovarismicroTube with the E220 system (Covaris, Inc., Woburn, MA, USA). The fragment size distribution was checked using Agilent Bioanalyzer (Agilent Technologies, Santa Clara, CA) with High Sensitivity DNA Kit (Agilent Technologies) according to the manufacturers instructions. The resulting fragmented DNA was cleaned up using HighPrep beads (MagBio Genomics, Inc, Gaithersburg, Maryland). These fragments were subjected to end-repair, A-tailing, and ligation of the Illumina multiplexing adaptors using the NEXTFlex DNA Sequencing kit as per the manufacturers instruction. The resulting ligated DNA was cleaned up using HighPrep beads (MagBio Genomics, Inc, Gaithersburg, Maryland)and size selected (400600bp) on 2% low melting agarose gel and cleaned using MinElute column (QIAGEN, India). These adapter ligated fragments were subjected to 10 rounds of PCR (denaturation at 98C for 2 min, cycling (98C for 30S, 65C for 30S and 72C for 1 min) and final extension at 72C for 5 min) using primers provided in the NEXTFlex DNA Sequencing kit. The PCR products were purified using HighPrep beads. Quantification and size distribution of the prepared library was determined using Qubitflourometer and the Agilent High Sensitivity DNA Kit (Agilent Technologies)