25-SGIA-Center#2
Total mRNA was isolated from 40 frozen cell aliquots received from the six different centers. Total RNA including miRNA was isolated on Qiacube Connect (QIAGEN, Germantown, MD, United States) using miRNeasy Tissue/Cells Advanced Mini kit (Cat # 217604, QIAGEN) according to the manufacturers instructions. Concentration and quality were measured by Nanodrop and 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, United States). cDNA libraries were generated using TruSeq Stranded Total RNA kit (Cat # 20020598, Illumina, San Diego, CA, United States) according to its protocol and sequenced on Illumina NextSeq 550 platform (Illumina, San Diego, CA, United States).