PromethION sequencing; Chromosome-length, trio-binned, diploid genome assembly of the naked mole-rat (Heterocephalus glaber) by integrating Nanopore long-read genome sequencing, 10X short read genome sequencing, and Hi-C sequencing - ONT Nanopore sequencing reads
Chromosome-length, trio-binned, diploid genome assembly of the naked mole-rat (Heterocephalus glaber) by integrating Nanopore long-read genome sequencing, 10X short read genome sequencing, and Hi-C sequencing - ONT Nanopore sequencing reads
Subordinate male (wild-type) naked mole-rat between 5 years of age were anesthetized with isoflurane and rapidly decapitated. One gonad was removed and flash frozen in isopentane on dry ice. Tissue for DNA extraction was homogenized in cell lysis buffer [10mM TRIS(pH8.0), 25mM EDTA, 100mM NaCl, 0.5% SDS, RNase A] and digested with proteinase K overnight at 37C. The resulting lysate was extracted twice with TRIS saturated phenol (pH8.0) followed by a single extraction with chloroform. DNA was precipitated from solution by adding 1/10th volume 3M Sodium Acetate and 2.5 volumes of 95% ethanol followed by gentle inversion. Precipitated DNA was spooled onto a glass rod and washed with excess 70% ethanol and allowed to partially air dry before being resuspended in an appropriate volume of EB [10mM TRIS (pH8.0)] DNA for library preparation was sheared to an average size of 10kb on a Covaris g-TUBE. Libraries were prepared with the Oxford Nanopore SQK-LSK109 library prep by ligation kit according to the manufacturerвЂ™s instructions. The resulting libraries were quantified by fluorometry using a Qubit High Sensitivity DNA quantification kit (ThermoFisher) and 100 fmol of each resulting library was used to load a Promethion flowcell.