GSM8383235: Ileum, NI-1; Mus musculus; RNA-Seq
Samples were defrosted on ice and mucosal scrapes were transferred into 2mL Bead Mill Tubes pre-filled with ceramic beads and 1mL cold TRIzolTM Reagent (Thermo Fisher). Samples were lysed with a bead beater then transferred to a new Tube and processed according to manufacturer's instructions. Briefly, the samples were incubated at RT then spun down at 4ºC and the supernatant was transferred to a new tube with chloroform to extract the RNA and spin down again. The RNA was precipitated by adding equal volumes of 70% EtOH and continuing the RNA purification process with the PicoPure RNA Isolation Kit (Thermo Fisher). RNA samples were normalized to 200 ng/µl with ddH2O. 1000 ng of RNA was then used as the starting material for cDNA synthesis and tagmentation using the NEBNext Poly A kit per the manufacturer's instructions.