GSM3832851: Normal rep1; Xenopus laevis; RNA-Seq
Total RNA was extracted from normal and EU-microinjected embryos using RNeasy Mini Kit (Qiagen). Nascent EU-RNA was biotinylated by incubating with 25 µM biotin-azide, 100 mM Tris-HCl pH 8.5, 1 mM CuSO4, and 100 mM ascorbic acid for 30 min at room temperature. The biotinylated EU-RNA was pulled down with magnetic streptavidin beads following the instructions provided by the Click-iTTM Nascent RNA Capture Kit (Invitrogen, C10365). cDNA libraries were prepared from purified EU-RNA (from EU-injected embryos) or total RNA (from normal embryos) following the instructions from the Ovation® Human FFPE RNA-Seq Library Systems (NuGEN) .