GSM9040575: SK-N-SH, NTC siRNA, replicate 2; Homo sapiens; RNA-Seq
Cells were either lysed directly in RLT buffer (Qiagen) and subjected to RNA extraction using TRIzol Reagent (Invitrogen), followed by RNA cleanup with the RNeasy Mini Kit (Qiagen), using the cleanup protocol provided by the manufacturer. SKNSH and SKNAS CRISPRi: Sequencing libraries were generated using the Illumina Stranded mRNA Prep kit. SKNSH siRNA: 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 using either dTTP for non strand specific library or dUTP for strand specific library. For the non strand specific library, it was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. For the strand specific library, it was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification.