PacBio genome sequence of Myxococcus xanthus DK101
The integrity of genomic DNA was examined on 1% agarose gels. DNA purity was checked using the NanoPhotometer spectrophotometer (IMPLEN, CA, USA). DNA concentration was measured using Qubit DNA Assay Kit in Qubit 2.0 Flurometer (Life Technologies, CA, USA). DNA size was examined on pulse field electrophoresis. The first step in the generation of a SMRTbell library was production of appropriately-sized double stranded DNA fragments generated by random shearing of DNA. The SMRTbell library was produced by ligating universal hairpin adapters onto double-stranded DNA fragments. The hairpin dimers formed during this process were removed at the end of the protocol using a magnetic bead purification step with size-selective conditions. Adapter dimers were also efficiently removed using PacBios MagBead kit. The final step of the protocol was to remove failed ligation products through the use of exonucleases. After the exonuclease and AMPure PB purification steps, sequencing primer was annealed to the SMRTbell templates, followed by binding of the sequence polymerase to the annealed templates. According to the effective concentration of library and output requirements, the sample was sequenced on PacBio Sequel platform.