GSM8257660: AL156_4; Homo sapiens; RNA-Seq
MDDCs present in the flowtrough  were lysed and total RNA was extracted using the NucleoSpin RNA Mini kit for RNA purification (Macherey-Nagel, 740955) according to the manufacturer's instructions Ribosomal RNA was removed from total RNA, followed by ethanol precipitation. After fragmentation, the first strand cDNA was synthesized using random hexamer primers. During the second strand cDNA synthesis, dUTPs were replaced with dTTPs in the reaction buffer. The directional library was ready after end repair, A-tailing, adapter ligation, size selection, enzyme digestion, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries have been pooled and sequenced on Illumina platforms, according to effective library concentration and data amount required.