GSM6030869: iPSC IEC, propionate 10 mM treatment, replicate 3; Homo sapiens; RNA-Seq
Cell were lysed with 350 µL RA1 lysis buffer for 5 min. The entire volume of RA1 solution was collected as cell lysate and total RNA. Total RNA from the iPSC derived IECs was extracted using the NucleoSpin RNA isolation kit (Macherey-Nagel, Duren, Germany) according to manufacturer's instructions. Quality of the RNA was checked using a NanoDrop One (ThermoFisher, Wilmington, Delaware, USA) and bioanalyzer (Agilent). Samples were sent to BGI (Hong Kong) for transcriptome sequencing using the DNBSEQ Technology Platform with 20M reads per sample. Library construction and RNA sequencing on DNBSEQ were conducted at Beijing Genomics Institute (BGI) for pair-end 150bp runs. At BGI, Genomic DNA was removed with two digestions using Amplification grade DNAse I (Invitrogen). The RNA was sheared and reverse transcribed using random primers to obtain cDNA, which was used for library construction. The library quality was determined by using Bioanalyzer 2100 (Agilent Technologies). Then, the library was used for sequencing with the sequencing platform BGISEQ-500 (BGI).