bulk RNA-seq of Merkel cell carcinoma
RNA sequencing libraries were prepared using the TruSeq Stranded mRNA LT Sample Prep Kit (Illumina). Briefly, 500 ng total RNA was used for poly(A) mRNA selection, fragmentation, and cDNA synthesis. The resulting cDNA was end-repaired, adenylated, and ligated using Illumina-specific adapters. PCR amplification was performed to enrich adapter-ligated fragments. The final library was purified and validated for size distribution using a Bioanalyzer (Agilent) and quantified by qPCR using a Kapa Library Quantification Kit (Kapa Biosystems). Libraries were pooled at equimolar concentrations and sequenced.