Castor canadensis Pacbio Whole Genome Sequencing
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.