GSM8324933: Non-Stim TA, Young Vehicle, 3; Mus musculus; RNA-Seq
RNA was isolated using Zymo RNA Miniprep kit with on-column Dnase treatment. 300 micrograms of total RNA was used for construction of sequencing libraries. A cDNA library was prepared for each sample using the the NEBNext PolyA mRNA and Ultra II Directional RNA library kit for Illumina. Libraries were barcoded for multiplexing with NEBNext Multiplex Oligos for Illumina unique dual index Primer Pairs. The multiplexed sample was quantified with qPCR (Kapa Biosystems) specific for Illumina sequencing primers and the average fragment size was determined with a Bioanalyzer high sensitivity DNA chip (Agilent Technologies). The pooled sample was sequenced, with paired-end, 50 base reads on an Illumina NovaSeq 6000 located in the Translational Science Laboratory at the College of Medicine, Florida State University.  The pooled data were demultiplexed into individual sample data.