WGS of Apis mellifera: drone
DNA was extracted from a single drone pupa from the DH4 line and sequenced using Pacific Biosciences SMRT sequencing (PacBio). Cells were lysed by grinding in liquid nitrogen followed by incubation at 55C in cell lysis solution and proteinase K. The solution was then treated with RNase A and proteins were precipitated using Protein Precipitation Solution (Qiagen) and centrifugation at 4C. DNA was precipitated from the resulting supernatant by adding isopropanol and ethanol and centrifugation at 4C. A 10kb PacBio library was generated and size-selection with 7.5kb cut-off was performed following the standard SMRT bell construction protocol. The library was sequenced on 29 SMRT cells of the RSII instrument using the P6-C4 chemistry, which generated 10.2 Gb of filtered data. N50 subread length was 8.8kb.