A targeted gene panel for circulating tumor DNA sequencing in neuroblastoma
The cfDNA isolation from blood samples was done according to the protocol of QIAamp Circulating Nucleic Acid Kit. The cfDNA quantity was assessed with dsDNA HS assay kit using Qubit Fluorometer 4.0 (Thermo Fisher Scientific). cfDNA quality was assessed with the Agilent High Sensitivity D1000 ScreenTape System (Agilent Technologies). Illumina TruSeq DNA indexed libraries were prepared using xGen Custom target capture Library Prep (IDT) incorporating unique molecular identifiers (UMIs). Libraries were quantified used Qubit Fluorometer 4.0 (Thermo Fisher Scientific). Pooled samples were sequenced on NextSeq 500 platform (Illumina) in a 2x150 paired-end format.