GSM6214089: DMS273-sgATG7-SQSTM1-3; Homo sapiens; RNA-Seq
Total RNA was extracted and purified with the use of RNeasy Mini Kit (Qiagen). 400 ng of total RNA was treated with NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB) to eliminate rRNA molecules and used for cDNA library preparation. cDNA synthesis followed by transcriptome library preparation were conducted by MGIEasy RNA Directional Library Prep Kit V2.0 (MGI tech), where dUTP were incorporated in the process of the 2nd strand cDNA synthesis, instead of dTTP, that block PCR amplification against the 2nd strand templates, enabling strand-specific transcriptome profiling. A 12-cycle PCR amplification was performed to increase library yield. Resulting transcriptome sequencing libraries were quantified by Qubit dsDNA HS Assay (Thermofisher) and their fragment size distribution was confirmed by TapeStation D1000 ScreenTape (Agilent).