GSM7401493: hIL-6_2; Homo sapiens; OTHER
Total RNA was isolated using the Quick-RNA Miniprep Kit (Zymo Research, Irvine, CA, USA) Total RNA was isolated from cells following incubation with 4-Thiouridine (s4U; 300 uM). Subsequently, the 4-thiol groups in the s4Uracil-labeled transcripts were alkylated with iodoacetamide (IAA). QuantSeq 3' mRNA-Seq (FWD) (Lexogen, Inc.) Illumina-compatible, indexed sequencing libraries were then prepared from the alkylated RNA samples (100 ng) according to the manufacturer's protocol for oligo(dT)-primed first strand cDNA synthesis, random-primed second strand synthesis, and library amplification.