GSM7994875: Hippocampus_5d; Mus musculus; RNA-Seq
Total RNA was extracted from the hippocampus and cortex of mice exposed to UPM (3, 5, and 7 days and 3, 8, 12, 16, and 20 weeks) using TRIzol reagent, according to the manufacturer's instructions (Invitrogen, Carlsbad, CA, USA). RNAs obtained from mice in the same experimental group was pooled. RNA quality was assessed using the Agilent TapeStation 4000 system (Agilent Technologies, Amstelveen, Netherlands). RNA was quantified  using a NanoDrop-2000 Spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). Only samples with RNA integrity > 7.0 were included in QuantSeq Analysis. A  cDNA library was constructed using the QuantSeq 3' mRNA-Seq Library Prep Kit (Lexogen, Inc., Austria) according to the manufacturer's instructions. Briefly , total RNA was hybridized with an oligo-dT primer containing an Illumina-compatible sequence at its 5' end to synthesize the first strand of cDNA by reverse transcription. The second strand of cDNA was synthesized using a random primer containing an Illumina-compatible linker sequence at its 5' end. The cDNA library was then amplified to add the complete adapter sequences required for cluster generation. Finally, high-throughput sequencing was performed by a single-end 75-base pair sequencing using a NextSeq 550 (Illumina, Inc., USA ).