title: GSM6792311: MM1S multiple myeloma cells co-cultured with HS5 stromal cells for 72 hours [RNA-Seq]; Homo sapiens; RNA-Seq
library_construction_protocol: Cells were counted and viability was assessed. CD138+ plasma cells were then purified using anti-CD138 microbeads (Miltenyi Biotech, Auburn, United States). CD138+ multiple myeloma cells were then aliquoted separately to perform RNA extraction. RNA was extracted from CD138+ multiple myeloma cells using the RNeasy MiniKit (Qiagen, Germantown, United States). RNA quantity was evaluated using the Qubit RNA Assay Kit (Life Technologies, Carlsbad, United States) and RNA quality was determined on the Bioanalyzer using the RNA Pico Kit (Agilent, Santa Clara, United States). We used at least 500 ng of total RNA for each sample. Next, library preparation was done with NEBNext Ultra RNA Library Prep Kit for Illumina (New England BioLabs, Ipswich, United States), was converted into a DNA library following the manufacturer's protocol. Library quantity was determined using the Qubit High Sensitivity DNA Kit and library size was determined using the Bioanalyzer High Sensitivity Chip Kit (Agilent). Finally, libraries were put through quantitative PCR using the Universal Library Quantification Kit for Illumina (Kapa Biosystems, Wilmington, United States) and run on the 7900HT Fast quantitative PCR machine (ABI, Grand Island, NY). Libraries passing quality control were diluted to 2 nM using sterile water, and then sequenced