GSM8900478: 48S Footprints of HEK293T WT replicate 1; Homo sapiens; RNA-Seq
HEK293T RPS26 p.R100_M115del homozygous cells were generated by CRISPR/CAS9. Translation initiation of HEK293T WT and RPS26del was analysed by 48S footprinting (TCP-seq). Cells were crosslinked by HCHO 0.3% for 5 min on ice. Reaction was stopped by 0.125 M Glycine pH 2. Cells were lysed in polysome lysis buffer (20 mM TRIS pH 7, 1.5 mM MgCl2, 100 mM KCl, 1.5% NP40). The lysate was digested by RNase I and loaded onto a sucrose gradient. The fractions containing the 40S were extracted and footprints were isolated by TRIzol. Extracted RNA were processed through library preparation according to the Ribo-seq protocol (Ingolia et al, 2012)
GEO
GSM8900478_r1