mRNA-Seq:Aged C57 male mice N-RAM dentate gyrus
The concentration and quality of purified mRNA was determined by Qubit 3.0 and Agilent 2100 Bioanalyzer, respectively. mRNA libraries were performed using VAHTSTM mRNA-seq V3 Library Prep Kit for Illumina and 236ng mRNA was used. First, mRNA was incubated with oligo dT beads to form mRNA-beads hybrid, which was subjected to fragmentation. Strand-specific library was prepared by inclusion of dUTP in cDNA synthesis and digestion with UDG. Size selection was performed to selectively enrich insertion size range from 350 bp to 450 bp. The sequencing was performed using NovaSeq 6000, by multiplexing samples with dual barcoding index and the sequencing depth was 50 million reads on average.