Whole genome sequencing of an Acinetobacter pitti strain found in bed bugs.
Bacterial cells were pelleted by centrifugation for 10 min at 5000 x g and the pellet was resuspended in 180 uL ATL buffer and used to extract the DNA using MagAttract HMW DNA Kit (Qiagen). The initial concentration of DNA was evaluated using the Qubit dsDNA HS Assay Kit (Life Technologies). 50 ng DNA was used to prepare the library using Illumina DNA Prep, (M) Tagmentation library preparation kit (Illumina) following the manufacturer's user guide. The sample underwent simultaneous fragmentation and addition of adapter sequences. These adapters are utilized during a limited-cycle PCR in which unique indices were added to the sample. Following the library preparation, the final concentration of the library (Table 1) was measured using the Qubit dsDNA HS Assay Kit (Life Technologies), and the average library size (Table 1) was determined using the Agilent 2100 Bioanalyzer (Agilent Technologies). The library was then pooled in equimolar ratios of 6pM, and sequenced paired end for 600 cycles using the MiSeq system (Illumina).