GSM9326448: Gln (0.20 mM), rep2; Homo sapiens; RNA-Seq
RNA was extracted using the Quick-RNA Microprep Kit (Cat# R1051, Zymo Research) according to the manufacturer's protocol. For RNAseq, libraries were prepared from 300 ng RNA with the KAPA RNA HyperPrep Kit with RiboErase (HMR) (Cat# KK8502, Kapa Biosystems), spiked with ERCC ExFold RNA Spike-In Mix 2 (Cat# 4456739, Invitrogen) at a final dilution of 1:1000. RNA was fragmented for 6 min at 94 °C to obtain a desired library insert size of 200-300 bp. Library concentrations were determined using the Qubit HS kit (Cat# Q32854, Invitrogen) and the library size distribution was determined using a High Sensititivy DNA analysis (Cat# 5067-4626, Agilent) on a Bioanalyzer 2100 (Agilent). Libraries were loaded on the NextSeq 2000 system (Illumina) with an average library size between 300-400 bp.