Nanopore DNA sequencing of Otocolobus manul: adult male whole blood
Genomic DNA was extracted from whole blood using a Quick DNA Miniprep Plus kit (Zymo Research) and seven library preps were performed using the SQK-LSK110 kit (Oxford Nanopore Technologies). The library prep protocol (GDE_9108_v110_revJ_10Nov2020) was modified to elute each library in 45 microliters of buffer. The eluent was then split into 15 microliter aliquots and each one was sequenced for 24 hours on a R9.4.1 minION flow cell using minKNOW software v22.05.5 and Guppy basecalling (v6.2.1). Nuclease flushes were performed between library reloads; three library aliquots were sequenced per flow cell for a total usage of seven flowcells and 21 sequencing days. Guppy basecalling was set to fast mode and super-accuracy basecalling (r941_min_sup_g507) was performed post-hoc.