GSM7848819: eIF1-2-KO_tachyzoite_RIBOseq_rep3; Toxoplasma gondii; OTHER
cytoplasmic lysate was collected and an aliquot was immediately stored in Trizol for RNA-seq samples.  The rest was digested with RNaseI and monosomes were collected by ultracentrifugation on a sucrose cushion. Ribosome-protected fragments (RFP) were purified after gel electrophoresis total RNA was fragmented by alkaline hydrolysis. Fragments were purfied by gel electrophoresis. fragments were dephosphorylated, ligated at 5' end with barcoded adapter, rRNA removal with eukaryotic Ribo-minus kit , phosphorylated, ligated at 3' end with SR-adapter, reverse transcribed & PCR with NEBnext barcoded adapters