GSM3067235: mP beads, serum starved MCF7 cells, rep 2; Homo sapiens; OTHER
RNA was extracted by acidic-phenol chloroform separation. The quality of the RNA samples was assessed using Agilent Bioanalyzer 2100 and Nanodrop ND-1000 Spectrophotometer (Thermo Scientific). Whole transcriptome library preparation was performed starting from 1 µg of total RNA with RIN ≥ 8. Following enrichment of poly-A containing mRNA molecules using poly-dT oligo-attached magnetic beads, all recovered RNA was processed using the Illumina TruSeq RNA Sample Preparation Kit (Illumina #FC-122-1001 #FC-122-1002) and the protocol v2 Rev. C. Completed libraries were evaluated by DNA quantification and Bioanalyzer analysis (mean fragment length = 274 bp), and then submitted for sequencing. RNA-seq was constructed with barcodes to allow multiplexing of 12 samples per lane. Sequencing was carried out by the NGS UniTrento Facility on Illumina HiSeq 2000 using the protocol HCS 1.5.15.1 in single reads.