GSM8001260: S16 control siRNA rep1; Rattus norvegicus; RNA-Seq
Schwann cells were homogenized in 500uL of Trizol (Thermo Fisher, #15596026) per well for 5 minutes on a rotator at low speed. Lysate was placed in RNAse free microcentrifuge tubes, 200uL of chloroform was added, and tubes were centrifuged at 12,000g for 15 minutes at 4°C. The aqueous layer was purified using the Zymo RNA Clean & Concentrator kit protocol (catalog #R1014, Zymo Research) at step 2. 1000 ng total RNA was sent to Novogene for library preparation after PolyA selection and Illumina sequencing (Novaseq 6000). 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.