PacBio RS II sequencing of Pseudomonas citronellolis str. P3B5
To isolate genomic DNA (gDNA), cells were grown overnight in terrific broth (12 g l-1 tryptone, 24 g l-1 yeast extract, 4 ml l-1glycerol, 100 ml l-1 0.17 M KH2PO4, 0.72 M K2HPO4) at 30 °C and 250 rpm. gDNA was extracted using the GenElute Bacterial DNA Kit (Sigma-Aldrich) according to the manufacturers recommendations. To concentrate the gDNA, the primary eluate was precipitated using 5M ammonium acetate and 100% ethanol followed by a washing step using 70% ethanol as described elsewhere {Sambrook, 1989 #1249}. The quality and quantity of the extracted DNA was evaluated by 1.0 % (w/v) agarose gel electrophoresis in 1xTBE buffer, by measuring absorption rations 260nm/280nm and 260nm/230nm, and additionally by performing a Qubit dsDNA GR assay (Life Technologies).  A 20kb PacBio library was created by Microsynth, Switzerland, and sequenced on the PacBio RS II platform by Microsynth, Switzerland.