GSM8078731: RNA_CM_WT_day3_rep2; Mus musculus; RNA-Seq
At day 3 or day 4 of CM differentiation, Srf-flex1 cells carrying H2B-mCherry (WT) or H2B-Venus-T2A-Cre (SRF KO) were trypsinized, resuspended in 3% BSA in PBS, and 40,000 cells (per replicate) expressing fluorescent reporter (mCherry or Venus) were sorted on a FACS Aria II (Becton Dickinson). Total RNA was extracted from 40,000 differentiated cells using Trizol reagent (ThermoFisher; 15596026) following the manufacturer's instructions and purified using the RNeasy Micro kit (Qiagen). Residual genomic DNA was digested on a column. The RNA was quantified using Qubit RNA HS assay (Life Technologies), and the integrity of the RNA was assessed using Bioanalyzer RNA pico chips (Agilent). Strand-specific RNA-seq libraries were generated from 100 ng of total RNA using the ScriptSeq v2 (Epicentre) low input library preparation kit according to manufacturer's instructions. The library was amplified using 15 PCR cycles. RNA-seq libraries were quantified using the Qubit HS DNA assay (Life Technologies) and the size distribution was assessed using the DNA HS Bioanalyzer chips (Agilent).