RNA-seq clone10 24h replicate 3
500ng of total-RNA was added to the ribosomal RNA (rRNA) depletion reaction using the NEBNext rRNA depletion kit (Human/mouse/rat). This step uses specific probes that bind to the rRNA in order to cleave it. rRNA-depleted RNA was then DNase treated and purified using Agencourt RNAClean XP beads (Beckman Coulter Inc, #66514 before fragmentation using random primers then first strand and second strand synthesis to create cDNA. cDNA was end-repaired before ligation of sequencing adapters, and adapter-ligated libraries were enriched by 10 cycles of PCR using NEBNext Multiplex Oligos for Illumina (96 Unique Dual Index Primer Pairs) (NEB, #E6440).