Illumina NovaSeq 6000 paired end sequencing; Simulated microgravity regulates the self-renewal and pluripotency of mouse and human embryonic stem cells - RNA-seq
Simulated microgravity regulates the self-renewal and pluripotency of mouse and human embryonic stem cells - RNA-seq
Total RNA was purified by RNeasy Minikit (Qiagen) according to the manufacturerâ€™s manual. RNA concentration was determined by Nanodrop, and 500 ng of total RNA was used for library construction using KAPA Stranded mRNA-Seq Kits. Sequencing was performed on Illumina HiSeq 2500 machines with 125 bp pair-end mode. Mouse E14 ESCs were cultured in DMEM (Hyclone) supplemented with 10% FCS (Gibco), 1xNEAA (Gibco), 1 mM sodium pyruvate (Gibco), 0.1 mM 2-mercaptoethanol (Sigma), 2 mM L-glutamine (Gibco), and LIF (Millipore) on 0.1% gelatin (Sigma) coated plates. Total RNA was extracted using the RNeasy Minikit (Qiagen) based on the manufacturer's instructions. Library was prepared using KAPA Stranded mRNA-Seq Kits based on the manufacturer's instructions.