GSM7401586: A375_DMSO-DMSO-3_R2; Homo sapiens; RNA-Seq
Cells were washed twice with ice cold PBS, then scrapped with 1ml of PBS. Then they were centrifuged (500g for 1min at 4°C) and slowly resuspended in 1ml of lysis buffer (10mM Tris-HCL pH7.5; 5mM MgCl2; 100 mM KCl; 1% Triton X-100), and incubated on ice for 10 minutes. After this, lysates were centrifuged (1000g for 10minutes at 4°C), and RNAs were extracted from cytoplasmic fractions using tri-reagent (Sigma, T9424) following the manufacturer's instructions. We prepared QuantSeq 3' mRNA-Seq Library Prep Kit (Lexogen, LEX113.96) libraries from RNA samples following the manufacturer's instructions. 500ng of RNA were spiked-in with 1µL of a 1:100 dilution of ERCC Spike-In Mix (ThermoFisher, 4456740) before the library preparation. Libraries were then quantified, pooled and sent for sequencing at Novogene (Novaseq 6000 1x150)