Libraries for whole genome sequencing (WGS) were prepared using the PCR-free
KAPA Hyper Prep Kit (Roche). For whole transcriptome sequencing, RNA samples were depleted of the ribosomal RNA using the
Ribo Zero Kit (Illumina) and library preparation was performed using the
TruSeq Stranded Total RNA Kit (Qiagen). For small RNA sequencing the
QIAseq miRNA Kit (Qiagen) was used All library preparation kits were used according to manufacturer’s instructions. Sequencing was performed on a
NovaSeq6000 system (Illumina).
For WGS, average coverage for tumor samples was ≥60X and ≥30X for normal samples with a total genomic coverage of ≥95%.
Whole transcriptome sequencing datasets have ≥100 million total reads with <20% of ribosomal origin and ≥20 million reads mapping to mRNAs according to Ensembl reference. Ribosomal depletion was performed to remove nuclear rRNA and mt-rRNA.
Han Y., Rovella V., Smirnov A., Buonomo O.C., Mauriello A., Perretta T., Shi Y., Woodmsith J., Bischof J., Melino G., Candi E, & Bernassola F. (2023). A BRCA2 germline mutation and high expression of immune checkpoints in a TNBC patient. Cell Death Discovery, 9, 370.