GSM8285737: HEK293T_PRCCTFE3_1; Homo sapiens; RNA-Seq
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. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount. Total mRNA isolation was performed using Trizol (TR 118) and isopropanol precipitation. RNA was then treated with DNAseI following the TurboDNAse kit instructions (AM1907 – ThermoFisher) and 1µg of RNA was reversed transcribed using SuperScript IV (18090050 – ThermoFisher) following manufacturer instructions.