GSM8987134: Resting fibroblasts, replicate 6; Mus musculus; RNA-Seq
Fibroblasts were isolated using the Magnetic Activated Cell Separation (MACS) technique using the Tumor-Associated Fibroblast Isolation Kit (mouse) from Miltenyi Biotec, Germany as per the manufacturer's instructions. RNA was extracted using the Zymo Research Quick-RNA Microprep Kit, as per the manufacturer's instructions. Quality of extracted RNA was assessed using the Agilent Bioanalyzer  RNA pico chips, and high quality RNA (RIN > 8.5) was taken forward into RNA-Seq library preparation. Firstly, oligo(dT)-primed, full-length cDNA was synthesised and amplified from total RNA input using the SMART-Seq mRNA kits (Takara Bio), as per the manufacturer's guidelines. cDNA was quantified using the Agilent Bioanalyzer HS DNA chips, and ~5 ng cDNA was taken into library preparation using the NEBNext Ultra II FS library preparation kits for Illumina (New England Biolabs).