GSM4729188: Tm4_polyH_2; Mus musculus; RNA-Seq
Total RNAs were isolated from 0.01 g of liver using the RNeasy Mini Kit (Qiagen) according to the manufacturer's instruction. For the isolation of the polysomal RNAs, the 'heavy' polysomal fractions (more than 3 ribosomes) were pooled and RNAs were precipitated in three volumes of 100% ethanol overnight at -20°C. The following day, RNAs were extracted using the RNeasy Mini Kit (Qiagen) according to the manufacturer's instructions. To allow quantification of RNAs from polysomal fractions, 200 ng of Luciferase RNA was spiked in to every fraction prior to precipitation when indicated (Luciferase control RNA, Promega). 2 μg and 0.5 μg of total and polysomal RNAs, respectively, were used to prepare sequencing libraries using the TruSeq Stranded mRNA Library Prep Kit (Illumina) according to the manufacturer's instructions.