GSM6617353: organoid_ctrl_sac_rpf_rep1; Homo sapiens; OTHER
For [Dataset 1] and [Dataset 2], CHX (cycloheximide, Milliporesigma C7698-1G) was added to a final concentration of 100 μg/ml to the medium and incubated for 7 minutes. Then spin down the cells, wash once with cold DPBS with 100 μg/ml CHX and spin down again, flash-freeze with LN2 and store at -80C. RPFs were prepared according to the manufacturer's instructions with riboLace kit (IMMAGINA, RL001). Total RNAs were extracted with standard TRIzol protocol. For [Dataset 1] and [Dataset 2], RNA libraries for input samples were constructed by SMARTer Stranded Total RNA-Seq Kit v2 (TaKaRa, 634417) with total RNA as input. RNA libraries for RPFs were constructed with NEBNext® Small RNA Library Prep Set for Illumina® (NEB, E7330L) with end-repaired RPFs extracted as inputs.