title: GSM5235810: CML cells from PDX mouse treated with ISRIB – rep2; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted using TRI Reagent (Sigma #T9424) or by Renozol (Genoplast #BMGPB1100-2) followed by Total RNA Mini column purification kit (A&A Biotechnology #031-100). 2 µg of RNA was subjected to reverse transcription using M-MLV enzyme (Promega #M1705), dNTP mix 100 mM each (BLIRT #RP65) and oligo (dT)18 primers (Bioline #BIO-38029). The RT-qPCR reaction was performed using SensiFAST SYBR Hi-ROX Kit (Bioline #BIO-92020) on the StepOnePlus™ platform (Thermo Fisher Scientific) according to MIQE guideline. Primers sequences are listed in the Supplementary Information. The comparative 2-ΔΔCt method was used to determine the relative mRNA level using StepOnePlus software. 18SrRNA was used as a reference control. The library was prepared using NEB Next Ultra II Directional RNA library Prep kit for Illumina (#E7335S/L).