RNA-Seq of mus musculus: midbrain
Total RNA was extracted from tissues using TRIzol (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA) and purified through chloroform and isopropanol precipitation. RNA quality and quantity were assessed by Agilent 2116 Bioanalyzer. Then, total RNA samples were isolated using oligo(dT)-attached magnetic beads. The isolated mRNA was fragmented and converted into cDNA through a two-step synthesis process involving first and second-strand reactions. The cDNA underwent end repair, the addition of a single 'A' nucleotide, and adaptor ligation. The library was amplified via PCR and quality controlled. The library products were then denatured and circularized. Single-stranded circular DNA molecules were replicated into DNA nanoballs (DNBs) via rolling circle amplification.