GSM8684493: 20670X4 Degs1fl/fl Sham Rep 4; Mus musculus; RNA-Seq
RNA was isolated from flash-frozen kidney cortex homogenized in 350 uL of TRIzol (Invitrogen, Cat# 15596026) using the Direct-zol RNA MiniPrep Kit (Zymo Research, Cat#R2052). RNA concentration and purity was determined with a NanoDrop 2000 Spectrophotometer (Thermo Scientific, Cat#ND2000). RNA concentration was measured with a Qubit RNA BR Assay Kit (Fisher Scientific, Cat#Q10211).  RNA quality was evaluated with an Agilent Technologies RNA ScreenTape Assay (Cat#5067-5576 and 5067-5577). Total RNA samples (5-500 ng) were hybridized with NEBNext rRNA Depletion Kit v2 (Human, Mouse, Rat) (New England Biolabs, Cat#E7400) to substantially diminish rRNA from the samples.  Stranded RNA sequencing libraries were prepared using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New Englan Biolabs, Cat#E7760L).  Purified libraries were qualified on an Agilent Technologies 4150 TapeStation using a D1000 ScreenTape assay (Agilent, Cat# 5067-5582 and 5067-5583).  The molarity of adapter-modified molecules was defined by quantitative PCR using the Kapa Library Quant Kit (Kapa Biosystems, Cat#KK4824).  Individual libraries were normalized and pooled in preparation for Illumina sequence analysis.