GSM5949596: G317_1_t8_exp; Mus musculus; RNA-Seq
Animal was sacrificed, decapitated and brain was removed from skull. Hippocampus isolation was done  in PBS, right and left halves of hippocampus were flash frozen.Total RNA isolation waswere carried out from the leftfrom left hippocampus with Trizol reagent (15596018, Ambion, Carlsbad, CA, USA) as described in the vendor's manual. The quality of RNA was assessed on an Agilent Bioanalyzer and confirmed fof their samples with RNA integrity numbers were between greater than 7.9. For mRNA-seq, the libraries were prepared using the Illumina® Stranded mRNA Prep, Ligation kit (Illumina) as per the manufacturer's instructions. Briefly, polyA RNA was purified from 200 ng of total RNA using oligo (dT) beads. The extracted mRNA fraction was subjected to fragmentation, reverse transcription, end repair, 3'– end adenylation, and adaptor ligation, followed by PCR amplification and SPRI bead purification (Beckman Coulter). The unique dual index sequences (IDT® for Illumina® RNA UD Indexes Set A, Ligation, Illumina) were incorporated in the adaptors for multiplexed high-throughput sequencing. The final product was assessed for its size distribution and concentration using BioAnalyzer High Sensitivity DNA Kit (Agilent Technologies). For microRNA-seq, the libraries were prepared using the NEXTflexTM Small RNA Library Prep Kit v3 (BioO Scientific) as per the manufacturer's instructions. The unique barcode sequences were incorporated in the adaptors for multiplexed high-throughput sequencing. The final product was assessed for its size distribution and concentration using BioAnalyzer High Sensitivity DNA Kit (Agilent Technologies). The denatured libraries were loaded onto an S1 flow cell on a NovaSeq 6000 or a Rapid flow cell on a HiSeq2500 (Illumina) and run for 2X53 cycles (for mRNA-seq) or for 1X49-65 cycles (for microRNA-seq) according to the manufacturer's instructions.