GSM9556300: 4T1 wild type with cytokines IFNg, IL1b (20ug/ml), biol rep 3; Mus musculus; RNA-Seq
1-1.5 million cells were grown under eight conditions in triplicate, pelleted, flash frozen, and stored at -80 °C.  Total RNA was extracted from thawed cell pellets with the PureLink RNA Mini Kit (Invitrogen).  RNA molecules were quantified on a Qubit 4 fluorometer using Qubit RNA Broad Range reagents (Invitrogen).  RNA quality was assessed on a Bioanalyzer 2100 using Agilent RNA 6000 Nano reagents (Agilent). RNA libraries were prepared with a Watchmaker mRNA Library Prep Kit (Watchmaker Genomics) using 1000 ng total RNA input and indexed for multiplexing with xGen Stubby Adapter and UDI Primers for Element (IDT).  Libraries were quantified on a Qubit 4 fluorometer using Qubit 1X dsDNA HS reagents (Agilent).  Library size distributions were assessed on an AATI Fragment Analyzer using the HS NGS Fragment Kit (Agilent).  The libraries were pooled to equimolar concentration and a final round of QC was performed on the pool, as described for the individual libraries.  The final pool was sequenced on an AVITI24 instrument using an AVITI 2x75 Cloudbreak Freestyle High Output Sequencing Kit (Element Biosciences).