GSM8389198: PyMT, Vehicle, replicate 1; Mus musculus; RNA-Seq
RNA extraction was carried out with RNeasy Plus Mini Kit (Qiagen).Total RNA was evaluated with Agilent Tapestation 4150 system and RNA ScreenTape (Agilent Technologies). All samples had an RNA Integrity Number (RIN) of 9 or higher. Samples were quantitated with the Qubit RNA HS Assay Kit. Total RNA input was normalized to 500 ng. mRNA was enriched using NEBNext Poly(A) mRNA Magnetic Isolation Module (New England BioLabs) and converted into an Illumina library using NEBNext Ultra II Directional RNA Library Prep with Sample Purification Beads (New England BioLabs). Indexed libraries were quality assessed using Qubit dsDNA HS Assay Kit (Invitrogen) and Tapestation 4150 DNA HS 1000 Kit (Agilent Technologies). The individual libraries were normalized, and equal molar amounts were multiplexed. Molar concentration of the multiplex pool was determined with KAPA Library Quantification Kits for Illumina.