title: MiSeq sequences of Microbacterium chocolatum GAI20246-6
design: Bacterial high molecular weight DNA was extracted using an adapted CTAB/chloroform-based protocol. In short, cell pellets of stationary growing bacteria were mixed with CTAB buffer, RNase A (10 mg/mL) and RNase T1 (1000U/uL). Cells were then incubated 20 min at room temperature followed by 30 min at 60C. An equal volume of chloroform:isoamylalcohol was added to the lysed cells and samples were centrifuged 20 min at 10,000xg at room temperature. Aqueous phase was recovered, mixed with equal volume of isopropanol and incubated overnight at -20C. HMW DNA was pelleted for 20 min at 10,000xg at 4C, washed twice with cold 70% ethanol and resuspended in 30 uL of low EDTA TE (pH 8.0). Quality of resulting DNA was analyzed on an Agilents 2200 TapeStation. Fragmentation was performed on a Covaris S220 instrument and libraries were built with the Accel-NGS 2S PCR-free DNA library kit (Swift Biosciences, discontinued). Final libraries were quantified using Picogreen standard curve and quality checked on the Agilents 2200 TapeStation. Sequencing was conducted on a MiSeq Illumina sequencer (paired-end 300 bp) at the J. Craig Venter Institute Sequencing Core (La Jolla, CA, USA).