GSM5651842: LIV-3-NC; Homo sapiens; RNA-Seq
Organoids cultured in either expansion or differentiation media were harvested into Trizol to isolate RNA. Total RNA was isolated from organoids and liver tissue using miRNeasy mini Kit, according to the manufacturer's protocol (QIAGEN, Valencia, CA). The samples were checked for quality and quantity using the Nanodrop 1000 spectrophotometer, Qubit fluorometer (Thermo Fisher Scientific, Waltham, MA), and Agilent 2100 Bioanalyzer (Agilent, Santa Clara, CA). Total RNA was reverse transcribed using Illumina's TruSeq RNA library preparation kit V2. Each sample was fitted with one of the 24 adapters containing a different 6 base molecular barcode for high level multiplexing. After 12 cycles of PCR amplification, completed libraries were sequenced on an Illumina instrument.