title: GSM3317100: WT cell line 26 SB; Mus musculus; RNA-Seq
library_construction_protocol: Cells were harvested from dishes using the RNeasy Plus Mini Kit (#74134, Qiagen) Briefly, 600 ul Buffer RLT was added to each plate, cells were scraped and transfered to a microfuge tube (RNase free). Samples were homogenized using the Qiashredder (79654, Qiagen) and transfered to -80C. The following day, RNA extraction was performed according to manufacturers instructions. RNA integrity was checked using the bioanalyzer and 1ug total RNA in up to 50ul, RIN>8, dissolved in RNase-, DNase-, protease-free water was sent to GATC Biotech for sequencing. pSmad levels were checked in protein extracts using SDS-PAGE and Immunoblotting to confirm activin stimulation RNA libraries were prepared by GATC Biotech for sequencing using standard Illumina protocols as part of their InView Transcriptome Explore package (https://www.eurofinsgenomics.eu/en/next-generation-sequencing/ngs-built-for-you/inview-transcriptome/inview-transcriptome-explore/). Briefly, mRNA is poly-(A) enriched and subsequently fragmented. cDNA synthesis is performed with random hexamer priming. 50 bp single reads with 30 million single reads guaranteed.
attributes.tag: GEO Accession
attributes.value: GSM3317100