Patient 22 : diagnosis
Blood was collected from each patient at three time points. At each collection, 8 mL of whole blood was drawn into a cell-free DNA BCT tube, stored at room temperature for no more than 7 days, and centrifuged at 1,900 x g for 10 min. The plasma supernatant was stored at -80 degrees C until cfDNA extraction using the QIAamp Circulating Nucleic Acid Kit. Extracted cfDNA was eluted in 30 uL and quantified using the Qubit dsDNA HS Assay Kit and Qubit 3.0 Fluorometer (Thermo Fisher Scientific, Waltham, MA, USA). Samples were then shipped to Macrogen Inc. (Seoul, South Korea) for lcWGS, where DNA integrity was verified using TapeStation cfDNA Screen Tape and libraries were prepared using the TruSeq Nano DNA kit (insert size: 350 bp) and sequenced on the NovaSeq 6000 platform with 100 bp paired-end reads.