NSUN2 shRNA knock down or control shRNA in 2D skin culture under differentiation conditions, using primary neonatal keratinocytes. This Ribo-seq data was generated in order to establish translation efficiency effects of NSUN2 knock-down.
NSUN2 shRNA knock down or control shRNA in 2D skin culture under differentiation conditions, using primary neonatal keratinocytes. This Ribo-seq data was generated in order to establish translation efficiency effects of NSUN2 knock-down. Libraries were prepared with a custom Ribo-seq protocol. Cells were harvested, lysed on ice with lysis buffer (containing 100 µg/ml cycloheximide and Turbo DNase I), and centrifuged. RNase I (Invitrogen, AM2294) was added to cell lysate and RNA was digested before T4 PNK labeling, and excision of 25-45 nt RNA fragments a 15% TBE-urea gel (EC6885BOX) run at 55C. Finally, gel extracted RNA was prepared as a library with the NEBNext Small RNA Library Prep kit (E7330S).