Ribo-seq of mus musculus: STHdh Q111Q111 striatal neurons
Following cell lysis, samples were digested with Rnase I and 80S monosomes were isolated by ultracentrifugation on sucrose gradients, followed by RNA extraction. RNA fragments in the range of 16-34 nucleotides were gel purified. After dephosphorylation, Univeral miRNA cloning linker (NEB) was ligated to the 3' end of RNA footprints, which were reverse transcribed into cDNA and linearized. Depletion of rRNA was performed using the Ribo-Zero Gold kit (Illumina) prior to linker ligation. Libraries were barcoded during PCR amplification and sent for sequencing.