Ribo-Seq of Miwi[+/RK]; replicate 3
Testis lysates were treated with RNAse I and subjected to monosome isolation with S400 centrifugation. Ribosome footprints were purified with 15% UREA-PAGE and further processed into libraries using the SMARTer smRNA kit for Illumina (Takara). The ribosome protected fragments were poly-adenylated and reverse-transcribed with an oligo(dT) primer and a template-switching oligo. Libraries were amplified by 12 PCR cycles.