GSM3145183: Isoxaflutole_0.1uM (RM-Sample-15); Homo sapiens; RNA-Seq
RNA extraction was performed using the Qiagen's RNeasy kit according to the manufacturer's instructions. A 100ng aliquot of total RNA from each sample was used to prepare total RNA libraries using the KAPA Stranded RNA-Seq Kit with RiboErase (KAPA Biosystems, Massachusetts, USA), and samples were randomised before preparation. Fragmentation prior to first strand cDNA synthesis was carried out using incubation conditions recommended by the manufacturer for intact RNA samples (94°C for 6 minutes). Polymerase chain reaction (PCR) was performed for 14 cycles for final library amplification. Resulting libraries were quantified using the Qubit 2.0 spectrophotometer (Life Technologies, California, USA) and average fragment size assessed using the Agilent 2200 Tapestation (Agilent Technologies UK Limited). The transcriptome of HepaRG cells exposed to glyphosate was sequenced employing the same strategy, except that the libraries were prepared as previously described (Mesnage et al., 2018). A total of three separate sequencing pools were created using equimolar quantities of each sample with compatible indexes; two with 17 samples each, and one with 16 samples. Paired-end reads of 75bp were generated for each library using the Illumina NextSeq®500 in conjunction with the NextSeq®500 v2 High-output 150-cycle kit (Illumina Inc., Cambridge, UK).
GEO Accession
GSM3145183