Illumina TruSeq DNA library from standard dark brown Neovison vison DNA (mink_3-247) sequenced on Illumina HiSeq 2500 with TruSeq SBS Kit v3
Genomic DNA from mink tissues was extracted using QIAGEN Mini Spin Columns, following manufacturer protocol (QIAGEN, Germany). Library preparation was performed with the TruSeq DNA HT Sample Prep Kit (Illumina, USA), with slight modifications to manufacturer protocol. In brief, between 500 ng and 1000 ng of genomic DNA was fragmented to a mean target size of approximately 300-400 bp using sonication in a Covaris S2 (Covaris, USA). Fragmented DNA was end-repaired, A-tailed, and indexed using TruSeq Illumina adapters with overhang-T followed by DNA purification from the reaction mixture with MinElute PCR Purification and QIAquick PCR Purification Kits (QIAGEN). Purified DNA with ligated adapters was used for size selection in 2% agarose SizeSelect E-Gel (Invitrogene, USA). Fragments of 400-500 bp were then isolated from agarose with Gel Extraction Kit (QIAGEN) and used for library amplification. Enriched libraries were purified using QIAquick PCR Purification Kits (QIAGEN), followed by 2% agarose SizeSelect E-Gel (Invitrogene, USA) electrophoresis to remove primer dimers. Library validation was performed with an Agilent 2100 Bioanalyzer with DNA High Sensitivity chip (Agilent, USA), and quantified with qPCR using a KAPA Library Quantification Illumina Kit protocol (KAPA Biosystems, USA).