Pacbio of [Candida] glabrata ATCC2001(CBS138):Pacbio sequencing of Candida grabrata
Genomic DNA of strain ATCC2001 (gift of Cecile Fairhead, Pasteur Institute) was made by preparation of spheroplasts, followed by lysis and spooling of high molecular weight genomic DNA after ethanol precipitation. The sequencing library was prepared using DNA Template Prep kit v.2 (3-10kb) following the PacBio shared protocol guidelines for preparing size-selected ~20kb SMARTbell templates.  Briefly, 7.5 microgam of genomic DNA was diluted to 150 ul and sheared using Covaris G-tube by centrifugation in an Eppendorf 5424 microcentrifuge at 4600 rpm for 60 seconds.  Sheared DNA was then purified using AMPure beads and quality controlled for concentration and size.  5 microgram of sheared DNA was then used for DNA damage repair and end repair.  End repaired DNA was purified using AMPure beads and ligated to SMRTbell adapter via blunt end ligation and exonuclease III/VII treatment.  The exonuclease treated SMRTbell template was purified again using AMPure beads and quality controlled for yield and library size.  The purified SMRTbell library was size selected on Blue Pippin with a size cut off of 5kb.  The size selected library was then sequenced on PacBio RS II using PacBio P4/C2 chemistry.
SUB6698559
m140326_073259_42159