GSM8084823: Igf2bp3+/-, replicate 3, RNC-seq; Mus musculus; RNA-Seq
Samples were dissolved in 1 ml lysis buffer and incubated on ice for 10 mins. After centrifugation at 12,000 g for 15 mins at 4°C, the supernatant was collected, layered onto a 5% to 50% sucrose gradient. Samples were peak fractionated by A260 absorption using the gradient station. mRNAs associated with 4 or more ribosomes were collected as polysome fractions and subjected to RNA extraction with TRIzol reagent. RNAs were purified by poly(A) selection and used for library construction using NEBNext ultra II directional RNA library prep kit for Illumina (NEB) according to the manufacturer's protocols. The libraries were sequenced on a Hi-seq X10 instrument.