GSM8455103: Lu_4w_RNA_XistBxC_-+_XX_1; Mus musculus; RNA-Seq
Frozen organs of 4w-, 9w-, 1.5y-week old mice were homogenized in appropriate amounts of TRIzol reagent (1 ml TRIzol reagent per 50-100 mg tissue) using the GentleMACSTM Dissociator RNA_02_01 program. 1 ml of homogenized organs was used for RNA isolation. Frozen organs of E14.5 embryos were thawed in 500 µl TRIzol reagent. Embryonic organs were homogenized using a pellet pestle. Another 500 µl TRIzol reagent was added, and RNA isolation was performed for all time points according to manufacturer's instructions (Invitrogen, TRIzol Reagent, Cat. #15596018). RNA concentration was measured by the NanoDrop Microvolume Spectrophotometer. 100 ng of RNA were used to generate poly-A captured RNA-seq libraries using lllumina's Stranded mRNA Prep, Ligation kit according to manufacturer's instructions. An eighth of reaction volume per sample was pipetted by the MANTIS liquid handler. Concentration and fragment distribution of completed RNA-seq libraries was measured on Agilent's TapeStation System.