Whole genome sequencing of sample collected from patient MEA054 at time of diagnosis timepoint.
PCR-amplified WGS libraries were prepared in-house from samples with less than 100 ng of genomic DNA using the NEBNext Ultra II FS DNA Library Prep Kit (E7805L). Enzymatic fragmentation was performed for 8 minutes to achieve a target insert size of 450 bp. PCR cycle number was adjusted per manufacturer recommendations based on input DNA mass to balance library yield against amplification-induced artifacts. NEBNext Multiplex Oligos for Illumina Unique Dual Index UMI Adaptors DNA Set 1 (E7395S) were used as dual-index adapters. Library size was assessed using an Agilent 4200 TapeStation System with High Sensitivity D1000 ScreenTape; concentration was measured by qPCR with KAPA Library Quantification Kits (KK4824) on a QuantStudio 5 instrument.