GSM8181848: WT_MII_2; Mus musculus; OTHER
Ribo-lite experiments were improved according to published procedure. Briefly, 500 oocytes were gathered in 1.5 ml Eppendorf LoBind microcentrifuge tubes and lysed with lab-made cell lysis buffer with CHX added. After digesting by RNase I, the pellet ribosome with ribosome-binding fragments (RPF) were centrifuged by Beckman Optima MAX-IP ultracentrifuge. Then the RPF were purified by electrophoresis, and the cDNA library were constructed with Dplex-Small-RNA-seq Kit (DIAGENODE, c05030001). The purified RPF were dephpsphorylated and tailed. After hybrided with RT primer and TSO, the RPF were RT into cDNA. Then the primer with barcode and index were added for ligation and amplification.