RNA-seq of Mus musculus: E18.5 pooled cerebellum
E18.5 whole cerebella were dissected and pooled. RNAs were extracted from the pooled cerebella with the mirVana miRNA Isolation Kit into two fractions: >200 nt and < 200 nt. Polyadenylated (PolyA+) RNA was purified from the > 200 nt fraction using the NEBNext Poly(A) mRNA Magnetic Isolation Module. Messenger RNA selection was performed using NEBNext Oligod(T)25 beads. First-strand cDNA was synthesized from heat-denatured purified mRNA using a Maxima H Minus First Strand cDNA Synthesis kit, followed by PCR Clean DX bead cleanup. The second strand cDNA was synthesized using the NEBNext Ultra Directional Second Strand cDNA Synthesis kit. cDNA was fragmented by Covaris LE220 sonication to achieve 250-300 bp average fragment lengths. The paired-end sequencing library was prepared following the BC Cancer Genome Sciences Centre strand-specific, plate-based library construction protocol on a Microlab NIMBUS robot.