GSM8595201: xpc-1 RNA-seq R2; Caenorhabditis elegans; RNA-Seq
L1 wildtype, csb-1, xpc-1 1 stage  C. elegans were collected in M9 and washed until the supernatant was clear, followed by incubation with TRIzol and chloroform. After centrifugation at 14,000g for 15 minutes at 4°C, the aqueous phase was mixed with an equal volume of isopropanol. Following centrifugation, the RNA pellet was washed several times and then resuspended in RNase-free water. Next steps, stranded and poly(A) enriched library preparation and sequencing were performed by Novogene. 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. Then the second strand cDNA was synthesized using dUTP, instead of dTTP. The directional library was ready after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification.