GSM1838391: RNAseq_Smad1_5_KO_3; Mus musculus; RNA-Seq
RNA was extracted using RNAeasy minikit (Qiagen). Libraries were prepared using the Illumina's TruSeq Stranded Total RNA kit protocol. In short, the first step involves the removal of ribosomal RNA (rRNA) using biotinylated, target-specific oligos combined with Ribo- Zero rRNA removal beads. The Ribo-Zero Human/Mouse/Rat kit depletes samples ofcytoplasmic rRNA and the Ribo-Zero Gold kit depletes samples of both cytoplasmic andmitochondrial rRNA. Ribo-Zero Globin kit depletes globin-encoding mRNA in addition tothe rRNA species targeted with Ribo-Zero Gold. Ribo-Zero Plant kit targets cytoplasmic andchloroplast rRNA. Following purification, the RNA is fragmented into small pieces usingdivalent cations under elevated temperature. The cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and random primers, followed by sec- ondstrand cDNA synthesis using DNA Polymerase I and RNase H. These cDNA fragmentsthen have the addition of a single 'A' base and subsequent ligation of the adapter. The products are purified and enriched with PCR to create the final cDNA library.
GEO Accession
GSM1838391