GSM5964165: H1299 with glucose and NH4Cl rep3; Homo sapiens; RNA-Seq
Total RNA from treated H1299 cells was extracted using the Total RNA Purification Plus kit (#48300, NORGEN BIOTEK CORP., Canada), followed by quality assessment by NanoDrop One (#70-105-8111, Thermo Fisher Scientific, USA). For mRNA library generation, 200 ng of total RNA was treated with NEBNext Poly mRNA Magnetic Isolation Module (#E7490L, New England Biolabs, USA) following the manufacturer's protocol. Subsequently, isolated mRNA was fragmented for 10 min. cDNA was synthesized and amplified for 12 PCR cycles using NEBNext Ultra II Directional (stranded) RNA Library Prep Kit for Illumina (#E7760L, New England Biolabs, USA) with NEBNext Multilex Oligos Indexes kit following the manufacturer's directions.