GSM4675878: EpCAM- Zeb1-/- 1 [NG_KO_1]; Mus musculus; RNA-Seq
Cells were resuspended in RLT Plus buffer (Qiagen) and stored at -80 degrees. Total RNA was isolated from EpCAM+ and EpCAM- Zeb1-/- LSK cells 14 days after the last dose of pIpC (n= 4 mice per genotype) using the RNeasy Mini Kit™ (Qiagen) according to the manufacturer's instructions. Total RNA stored in RNase-free water was amplified using SMART-Seq™ v4 Ultra™ Low Input RNA Kit for Sequencing (Takara Bio USA, Inc., Mountain View, USA) with the double strand cDNA (ds-cDNA) being synthesized. Then, ds-cDNA is purified with AMPure XP beads and quantified with Qubit (Life TechnologiesTM). 1 ng cDNA per sample was used as input to construct 250~300 bp insert cDNA libraries using NEBNext® UltraTM RNA Library Prep Kit (New England Biolabs, Ipswich, USA) following the manufacturer's recommendations. Libraries were sequenced by Novogene, as 150-bp paired-end reads on an Illumina Novaseq 6000 platform.