RNA-Seq of Homo sapiens: Melanoma metastatic tumor, Brain
Extraction of RNA from frozen melanoma metastatic tumor brain tissue was performed using the RNeasy Plus Mini Kit (QIAGEN, CAT # 74134) according to the manufacturer's protocol. Extracted total RNA quality and concentration were assessed on a 2100 Bioanalyzer (Agilent Technologies) and Qubit 2.0 (ThermoFisher Scientific). RNA-seq libraries were prepared using Illumina Stranded Total RNA Prep with Ribo-Zero Plus kit (Illumina, CAT # 20040529) and indexed using Illumina RNA UD Indexes Set A (Illumina, CAT # 20040553). All libraries were then pooled in equimolar ratio and sequenced on 1 lane of an Illumina NovaSeq 6000 S4 flow cell, generating 150bp PE reads.  Sequence data was demultiplexed and Fastq files generated using Illumina Bcl2Fastq conversion software.