GSM7050420: Brain, high PAM, Rep 3; Mus musculus; RNA-Seq
Total RNA was isolated from day 35 pulverized pup left brain hemispheres (≤ 100mg starting material) and livers (≤ 5 mg starting material) using the RNeasy Lipid Tissue Mini Kit (Qiagen; item no. 74804) and the RNeasy Micro Kit (Qiagen; item no. 74004), respectively, according to manufacturer instructions. For liver RNA, extraction was followed by a 3M sodium acetate (Sigma; item no. 567422) re-purification step to remove contamination. Libraries were prepared following the NEB NEBNext Ultra II Directional RNA Library Preparation protocol to generate stranded data. Briefly, 400 ng of total RNA was used as input material and enriched for poly-A mRNA using magnetic oligo d(T) beads, fragmented into the 200-300-bases range for 10 minutes at 94oC and converted to double stranded cDNA. Next, cDNA proceeded to library preparation and dual-index Illumina adapters were added through PCR.