title: Castor canadensis Pacbio Whole Genome Sequencing
design: PacBio SMRTbell libraries were constructed from purified genomic DNA from fresh beaver blood leukocytes and sequenced on the PacBio RSII SMRT platform.  The DNA shearing step was omitted since pulsed-field gel analysis showed that genomic DNA already had a broad size distribution with a peak mode length of ~40 kb, similar to the recommended size for library construction.  Libraries were prepared from 8 µg of genomic DNA using the SMRTbell Template Prep Kit 1.0 according to the manufacturer’s instructions for 20 kb insert templates.  SMRTbell libraries were size-selected using the BluePippin System (Sage Science, Beverly, MA, USA) with a lower cut-off of 15 kb.  Whole-genome SMRT sequencing used 80 flow cells, running the P6/C4 chemistry with a 6 hr movie-time, and used PacBio’s P-Filter Module protocol to process raw data to remove library adaptors and redundant or low-quality sequence reads.  Each flow cells produced an average of 1 Gb of filtered PacBio reads with average read length of ~13-14 kb.