WGA of Myxococcus xanthus natural isolates
A total of 10 g of gDNA were sheared to an average size distribution of 10 Kbp, using a gTube (Covaris , USA). A Bioanalyzer 2100 12K DNA Chip assay (Agilent, USA) was used to assess the fragment size distribution. A total of 5 g of sheared gDNA was DNA-damage repaired and end-repaired using polishing enzymes. A blunt-end ligation reaction followed by exonuclease treatment was performed to create the SMRT bell template. The library was quality inspected and quantified on the Agilent Bioanalyzer 12Kb DNA Chip and on a Qubit Fluorometer (Invitrogen, USA), respectively. A ready-to-use sequence SMRTbell-Polymerase Complex was created using the P4 DNA/Polymerase binding kit 2.0 (Pacific Biosciences, USA) according to the manufacturers instructions. The Pacific Biosciences RS2 instrument was programmed to load and sequence the samples on PacBio SMRT cells v3.0 (Pacific Biosciences, USA), taking one movie of 240 minutes each per SMRT cell. The MagBead (Pacific Biosciences, USA) loading method was chosen in order to improve the enrichment of longer fragments. After the run, a sequencing report was generated for every cell, via the SMRT portal, in order to assess the adapter dimer contamination, the sample loading efficiency, the obtained average read-length and the number of filtered sub-reads.