Illumina HiSeq 2000 paired end sequencing; RNA-Seq of Ras induced Epithelial to Mesenchymal Transition (EMT) in Madin-Darby Canine Kidney (MDCK) cells
RNA-Seq of Ras induced Epithelial to Mesenchymal Transition (EMT) in Madin-Darby Canine Kidney (MDCK) cells
MDCK and MDCK-Ras cells were cultivated in DMEM/F12 +10%FCS. Confluent cultures were used for the experiment. Starting with an amount of 15 µg total RNA poly(A)+ selected cDNA samples were generated following a protocol preserving the strand information based on the dUTP method [Mortazavi et al., Nature Methods 2008]. Total RNA was isolated from 4 biological replicates from each cell type (MDCK and MDCK-Ras) using peqGOLD TriFast (Peqlab) according to the manufacturer’s instructions. Quality and quantity of total RNA was checked on Agilent Bioanalyzer according to manufacturer’s instructions. Strand specific and indexed cDNA libraries with 200 – 700 bp insert size were generated for sequencing using the NEBNext® Ultra™ DNA Library Prep Kit for Illumina® (New England BioLabs).
INSTITUTE OF ANIMAL BREEDING & GENETICS, UNIVERSITY OF VETERINARY MEDICINE VIENNA
E-MTAB-3301:Mueller_Lab_Feb2015_MDCK_biorep4_techrep3
Institute of Animal Breeding & Genetics, University of Veterinary Medicine Vienna
E-MTAB-3301:Mueller_Lab_Feb2015_MDCK_biorep4_techrep3
Experimental Factor: phenotype
epithelial
LIBRARY_STRAND
first strand