GSM9235583: iNeurons - TBK1 KO (A)- biol rep 3; Homo sapiens; RNA-Seq
Each sample consisted of 4 million cells, with RNA extracted from four independent passages. The RNA was harvested using the RNeasy Plus Mini Kit (Qiagen) and the concentration for each hiPSC sample was standardized to 500 ng/µL.For differentiated iNeurons, RNA was harvested at D21 using the RNeasy Micro Kit (Qiagen) and RNA met a minimum concentration threshold of 150 ng/µL. . 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, followed by the second strand cDNA synthesis using either dUTP for directional library or dTTP for non-directional library.For the non-directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. For the directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification.The library was checked with Qubit and real-time PCR for quantificationand bioanalyzer for size distribution detection. Quantified libraries will bepooled and sequenced on Illumina platforms, according to effective libraryconcentration and data amount.