Whole-genome sequencing of Apis mellifera drones: Sample OU22
"Total DNA was extracted from the thorax of the drones or from whole larvae cut into small pieces with a scalpel. The fragments were first incubated 3 hours at 56o in 1 mL of a solution containing 4 M urea, 10 mM Tris-HCl pH 8, 300 mM NaCl, 1% SDS, 10 mM EDTA and 0.25 mg proteinase K, after which 0.25 mg proteinase K was added for an incubation over-night at 37oC. Four hundred uL of a saturated NaCl solution was added to the incubation, which was then gently mixed and centrifuged for 30 minutes at 15000 g. The supernatant was treated for 5 minutes at room temperature with RNAse (Qiagen) and then centrifuged again, after which the DNA in the supernatant was precipitated with absolute ethanol and re-suspended in 100 uL TE 10/0.1. Paired-end sequencing was performed on Illumina HiSeq 2000 and HiSeq 2500 platforms, with 20 samples per lane, following the manufacturer's protocols for library reparations."