GSM4697449: C1-L; Canis lupus familiaris; MBD-Seq
Tumor cells from cHGBL cases were collected via fine needle aspiration of the enlarged lymph nodes and preserved in RPMI medium until genomic DNA extraction. Whole blood was collected from each healthy dog and peripheral blood mononuclear cells were separated by Ficoll-Paque PLUS gradient centrifugation for genomic DNA extraction. Total genomic DNA was extracted from no more than 5 x 106 cells using Genomic DNA isolation Kit. A total amount of 800 pg captured DNA (meDNA captured sample) was taken for library preparation using QIAseq Ultralow Input Library Kit according to the manufacturer's protocol. Briefly, the double-stranded DNA was end-repaired and 3'end adenylated followed by adapter ligation and PCR amplification. The length of library fragments ranging from 200 bp and 1 kb were further selected using AMPure XP beads. The fragment size of the resulting library was validated using Agilent 2200 TapeStation-D1000 Screentape system. The library was quantitatively measured by Kapa Library Quantification Kit for Illumina platform and Qubit dsDNA HS Assay kit. The library was sequenced using a 150 bp paired-end protocol on an Illumina Novaseq.