RNA-seq of DHCR7 silenced
Total RNA from the lysed cells was extracted with Nucleospin RNA II Kit (Macherey-Nagel). Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. Following is the workflow of library construction