GSM7506276: DMSO 4sU rep2; Homo sapiens; RNA-Seq
RNA was extracted using TRIzol(Thermo) with the following modifications of the manufacturer protocol. After the transfer of the aqueous solution, 1% vol/vol 100 mM DTT was added to keep the RNA in an unoxidized state. After dissolving the RNA pellet in 1x PBS, 5 µg of RNA in a vloume of 90 µl were ued for Iodacetamide treatment.  90 µlof 20 mM Iodacetamide were added and incubated at 50°C for 15 min. The reaction was quenched by the addition of 20 μl of 1 M DTT, and the RNA was precipitated by NaAc/EtOH. The RNA was then used for QuantSeq library prep. 3'Seq libraries were generated using the QuantSeq 3'-mRNA-Seq Library Prep Kit FWD for Illumina (Lexogen) according to the manufacturer's protocol.