GSM1542247: TT-FHAgo2_nodox96h_rep2; Mus musculus; RNA-Seq
For ESC, strand-specific mRNA sequencing libraries were prepared using either the UTP method or Truseq sample preparation kit from Illumina. Muliplexing barcode sequences were incorporated during the PCR amplification. For MSC, libraries from polyA RNA were generated with the Illumina TruSeq kit. To estimate copies per cells, a separate RNA-seq biological replicate was performed for ES and MSCs with spike-in standard RNAs. Cells were counted before extracting total RNA. 1.5ng External RNA Controls Consortium ERCC (Life Technologies) mix 1 RNA spike-ins of known concentrations were added to the cellular RNA post oligo dT purification step, corresponding to ~1-5% of total poly A RNA input into Illumina sequencing prep. FPKM expression values were converted based on regression fit for the known concentration of standards, and this was related to cell counts by total RNA input into prep.