title: GSM8616975: Hippocampus, CLO-2; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA from the hippocampus of offspring mice was extracted using the NucleoSpin® Plus (Macherey-Nagel GmbH & Co., Düren, Germany). 300 ng of total RNA samples, which met the quality guideline, RIN: 8 or higher, were treated with NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB) to enrich poly-A mRNA and to remove rRNA molecules. cDNA synthesis followed by transcriptome library preparation were conducted by NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB), where dUTP were incorporated in the process of the 2nd strand cDNA synthesis, instead of dTTP, that block PCR amplification against the 2nd strand templates. This enables to maintain the strandness of RNA transcripts. A 10-cycle PCR amplification was performed to increase library yield and also to incorporate sample barcodes into the library fragments. Resulting transcriptome libraries were quantified by Qubit DNA Assay (Thermofisher) and their fragment size distribution was estimated by TapeStation D1000 ScreenTape (Agilent).