title: GSM1404609: d0 RNAseq; Mus musculus; RNA-Seq
library_construction_protocol: RNA-seq: Nuclear lysate was removed from CLIP protocol before sonication and treated with proteinase K and reprecipitated in the same way as CLIP RNA eluted from membrane. RNA was depleted of ribosomal RNA with RiboMinus Eukaryote System (Ambion A15026), treated with TURBO DNase, and cleaned up with RNeasy MinElute columns (Qiagen 74204). RNA was fragmented in Superscript III first strand synthesis buffer before cDNA synthesis. RNA-seq: Fragmented RNA was used for first strand cDNA synthesis with the addition of actinomycin D. Second strand cDNA synthesis was performed with NEBNext mRNA Second Strand Synthesis Module (E6111) in the presence of dUTP to preserve directionality. Double-stranded cDNA was then used to generate RNA-seq libraries using the NEBNext ChIP-Seq Library Prep Master Mix Set (E6240).
attributes.tag: GEO Accession
attributes.value: GSM1404609