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WGS of E. coli PB11
Genomic DNA libraries were constructed using the TruSeq DNA Sample prep kit (Illumina, San Diego, CA, USA). Genomic DNAs were quantified using the Qbit BR reagents (Qbit 2.0 FluorometerLife Technologies). Briefly, 1 ug of DNA in 130 ul was sheared into 300-bp fragments on a Covaris S2 focused ultrasonicator (Covaris Inc) using SonoLab software for 200 cycles/burst, 10% Duty factor, 4 Intensity, in screw-cap microtubes. After shearing, the DNA was blunt-ended, 3-end A-tailed and ligated to indexed adaptors. The adaptor-ligated genomic DNA was size selected with AMPure-beads using the gel-free protocol described in the TruSeq DNA Sample Prep manual. Size-selected DNA was amplified by PCR to selectively enrich for fragments that have adapters on both ends. Final amplified libraries were run on an Agilent bioanalyzer DNA 2100 (Agilent, Santa Clara, CA, USA) to determine the average fragment size and to confirm the presence of DNA of the expected size range.
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SUB6639232
150409_1