GSM3098347: 1009_young_F_pre bed rest; Homo sapiens; RNA-Seq
10-15mg of tissue was placed in Tri reagent LS (Molecular Research Center, Cincinnati, OH, USA) and disrupted via hand-held homogenizer (Bio-Gen PRO200; Pro Scientific, Oxford, CT, USA).  Chloroform was added to separate the RNA into an aqueous phase that was then precipitated using isopropanol.   Extracted RNA was then washed with ethanol and suspended in nuclease-free water with EDTA.  RNA integrity and quantity was determined using the EPOCH (Take3; BioTek) spectrophotometer. Total RNA samples (100-500 ng) were hybridized with Ribo-Zero Gold to substantially deplete cytoplasmic and mitochondrial rRNA from the samples.  Stranded RNA sequencing libraries were prepared as described using the Illumina TruSeq Stranded Total RNA Kit with Ribo-Zero Gold (RS-122-2301 and RS-122-2302).  Purified libraries were qualified on an Agilent Technologies 2200 TapeStation using a D1000 ScreenTape assay (cat# 5067-5582 and 5067-5583).  The molarity of adapter-modified molecules was defined by quantitative PCR using the Kapa Biosystems Kapa Library Quant Kit (cat#KK4824).  Individual libraries were normalized to 10 nM and equal volumes were pooled in preparation for Illumina sequence analysis.
GEO Accession
GSM3098347