RNA seq of MCF-7 cells extracted from the center of the micropatterns-2
RNA from the micro-dissected cells was extracted using the PicoPure RNA Isolation Kit (Arcturus) following manufacturers instructions. Concentrations of RNA samples were determined by absorbance on a NanoDrop One spectrophotometer and Qubit (ThermoFisher). RNA integrity was determined using an Agilent 2100 Bioanalyzer and RNA 6000 Nano Chip kit. Only samples with RNA Integrity Number (RIN) score greater than 7 were used. Following mRNA purification from total RNA using poly-A selection, a cDNA library was prepared using Illumina TruSeq Stranded mRNA Library Prep kit. The cDNA library quality was checked using the Agilent 2100 Bioanalyzer and D1000 DNA Chip kit. Samples were run on the Illumina NextSeq 500 platform using the High Output Sequencing Kit v2 (150 cycles, 2x75bp read length, 20 million reads per sample).