GSM9897996: MuSC RNA-seq III BMP6 3h biol rep 1; Mus musculus; RNA-Seq
Cells were washed, trypsinized, centrifuged, and frozen at -80 °C for later RNA extraction. RNA was extracted using the NucleoSpin RNA kit (Macherey-Nagel, 740955). RNA quantity and quality were assessed using a NanoDrop® 2000c spectrophotometer (Thermo Fisher Scientific) The construction of the eukaryotic strand-specific transcriptome library was performed according to the "DNBSEQ Eukaryotic Strand-specific Transcriptome Resequencing" protocol of the  Beijing Genetics Institute (BGI). Briefly, 500 ng total RNA was enriched for poly(A)-carrying molecules using oligo-dT beads. The RNA was then fragmented and the first cDNA strand was synthesized using random primers. Second strand cDNA was synthesized using dUTP instead of dTTP and the synthesized cDNA was subjected to end repair and 3' adenylation. Adaptors were ligated to the ends of these 3' adenylated cDNA fragments, which were then amplified by PCR. After quality control, the library was circularized and amplified to produce DNA nanoballs (DNBs).