BG2_Pacbio_denovo
Genomic DNA of strain BG2 was made by preparation of spheroplasts, followed by lysis and spooling of high molecular weight genomic DNA after ethanol precipitation. The genomic DNA of the four isolates were sheared to 30 kb with MegaRupter . Samples were diluted to under 50 ng/l per the MegaRupter protocol with TE.  Samples were then purified using a 1x AMPure bead cleanup and elutied in 62 l of EB.  Libraries were prepared using the standard PacBio 30kb protocol. Briefly,  after fragmentation DNA end repaired and A-tailed followed but SMRTbell hairpin ligation. Upon completion of the library, samples were then size selected on the Blue Pippin to the range of 20-50kb. The size selected library was then sequenced on PacBio RS II using PacBio P6/C4 chemistry.