GSM1885054: Flag-Lef1 plus Dox_3; Mus musculus; RNA-Seq
ES cells were havested on day 3 of culture plus/minus 24 treatment of doxycycline. Total RNA was extract using standard trizol-based extaction protocols. Total RNA was then treated with Turbo DNase (Ambion) to eliminate genomic DNA contamination. The libraries were prepared according to Illumina’s TruSeq RNA library protocol. We use between 100ng to 1ug of total RNA as the input to an mRNA capture with oligo-dT coated magnetic beads. The mRNA is fragmented, and then a random-primed cDNA synthesis is performed. The resulting double-strand cDNA is used as the input to a standard Illumina library prep with end-repair, adapter ligation and PCR amplification being performed to give you a library that would go to the sequencer.