title: GSM8688991: TFEB KO (clone #c20) DMSO biol rep 1; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was extracted from each well using 500 µL of Trizol followed by two sequential chloroform extractions followed by ethanol precipitation per the Trizol user guide (Thermo Fisher MAN0001271). 500ng total RNA was used as starting material for each sample and then depleted of rRNA using the Illumina rRNA depletion kit (NEB E6310). For Batch 1 samples, 1uL of 1:100 diluted ERCC RNA Spike-In Mix (Thermo #4456740) was first added to the 500ng starting total RNA before rRNA depletion and libraries were prepared for Illumina sequencing using the NEBNext Ultra II directional RNA library preparation kit for Illumina (E7760) per the manufacturer's protocol. For Batch 2 samples, libraries were prepared using the NEBNext Ultra II RNA Library Prep Kit for Illumina (E7770L) per the manufacturer's protocol.