ES_09_T_03 mRNA
Archived frozen tumor samples were weighed and up to 20-30mg were homogenized in RLT buffer, followed by extraction using the AllPrep DNA-RNA Mini Kit (Qiagen catalog 80204) according to the manufacturer's instructions. RNA was eluted in 13 uL nuclease-free water. After RiboGreen quantification and quality control by Agilent BioAnalyzer, 1 ug of total RNA with DV200 percentages varying from 78% to 100% underwent ribosomal depletion and library preparation using the TruSeq Stranded Total RNA LT Kit (Illumina catalog RS-122-1202) according to instructions provided by the manufacturer with 8 cycles of PCR. Samples were then barcoded and sequenced in PE150 mode with the NovaSeq 6000 S4 Reagent Kit (Illumina).