GSM4399896: control rep3_RNC; Mus musculus; RNA-Seq
Total RNA of a liver sample was extracted using the TRIzol reagent. RNC isolation was performed in accordance with a previously reported procedure with some modifications. Briefly, 90 mg liver tissue was treated with 200 μl lysis buffer (1% Triton X-100 in ribosome buffer [20 mM HEPES, 2 mM dithiothreitol, 15 mM MgCl2, 200 mM KCl and 100 µg/ml cycloheximide]) and was homogenized. An additional 700 μl lysis buffer was added to the samples, followed by incubation in an ice-bath for 30 min. After centrifuging at 16,200 x g for 20 min at 4°C, supernatants were added to the surface of 5 ml of sucrose buffer (30% sucrose in ribosome buffer), and the RNC complex was pelleted by ultra-centrifugation at 330,000 x g for 3 h at 4°C. RNC-RNA was extracted from the RNC complex using TRIzol. RNA libraries were prepared according to the protocol of the VAHTS mRNA-seq v.3 Library Prep Kit for Illumina (Vazyme Biotech Co., Ltd, Nanjing, Jiangsu, China).
GEO
GSM4399896