RNA was isolated from each sample using TriReagent® (MRC Inc., USA) according to manufacturer's recommendations. We had three technical replicates for each cell line (a, b and c, resulting in 15 samples in total). Genomic DNA was removed using RNeasy MinElute Cleanup kit (Qiagen, Germany). Quantity and quality were assessed in a Nanodrop ND-1000 machine (Thermo Scientific). The RNA integrity was assessed by gel electrophoresis and by an Experion system (BioRad Laboratories) using the ExperionTM RNA StdSens analysis kit (BioRad, Sweden). Average RNA quality indicator values (RQI) were 9.7 for the American mink ES cells, 9.6 for iPS cells and 9.3 for EF. This RNA was used for RNAseq library preparation and for cDNA synthesis, gene silencing analysis and qPCR.
One microgram of DNase I treated total RNA was used for cDNA synthesis [48 (link)]. cDNA synthesis was done in duplicates for each RNA sample, thus resulting in 30 samples. All samples were diluted eight times before using in qPCR.
One microgram of DNase I treated total RNA was used for cDNA synthesis [48 (link)]. cDNA synthesis was done in duplicates for each RNA sample, thus resulting in 30 samples. All samples were diluted eight times before using in qPCR.
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