GSM4975517: SMC 0hr replicate 3; Homo sapiens; RNA-Seq
Cultured hcSMCs were washed once with PBS and lysed directly with the lysis buffer in the flasks, and total RNA and DNA were extracted using RNA/DNA/Protein Purification Plus Kit (Norgen #47700). The DNA samples was quantified using Qubit® 2.0 Fluorometer (Life technologies). RNA-seq libraries were prepared with NEBNext Ultra™ Directional RNA Library Prep Kit for Illumina (NEB, E7420) and NEBNext rRNA Depletion Kit (E6310). PCR library enrichment was performed by 15 cycles of PCR with NEBNext Multiplex Oligos for Illumina (NEB #E7335). Quantification of library was performed using an Agilent 2100 Bioanalyzer and a Kapa Library Quantification Kit (Kapa Biosystems).