GSM7911900: MIN6 cells/25mM/RPF biol rep4 [RPF 8]; Mus musculus; OTHER
Ribosome protected footprints (RPFs) were obtained using RNase digestion and gel-extraction, as previously described (McGlincey et al. 2017) except that 1 U/20 million cells RNase 1 (Thermo Scientific) was used. Libraries were made in two batches (samples 1-8 in one batch; 9-16 in a second batch). For RPF library, rDNA depletion was performed using biotinylated rDNA sequences to remove rRNA contamination. Input libraries were generated using polyA enrichment, and Kapa stranded mRNA Hyper Prep (Illumina). RPF and RNA libraries were sequenced using Illumina NS500 single-end 75 bp reads. Ribosome profiling under low and high glucose conditions in MIN6 cells was designed to assay mRNA translation efficiency changes under different glucose conditions