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Tips for Working with RNA

  • When removing tissue or cells from the source (animal, tissue culture plates, etc.) all materials that come into contact with material should be sterile - RNA degrades quickly!
    • Gloves should be sterile and/or strayed with RNAse Decontamination spray or 70% EtOH before use and between samples
      • Avoid touching objects that contain RNAses (face, clothing, benchtop)-if this does occur, switch gloves before proceeding
    • All tools should be autoclaved and avoid touching unnecessary objects when not in use
      • This includes tissue culture plates, surgical tools, tubes for sample storage, etc.
      • Tools should be sterilized between samples by submerging in EtOH briefly
    • All buffers (PBS, tissue culture medium, etc.) should be cold and filtered before use
      • Keep cold in between uses
    • Wearing a face mask is also recommended before isolation procedure
    • Samples should be snap-frozen in liquid nitrogen if not immediately processed in tubes that are sterile
  • Keep material for RNA isolation cold AT ALL TIMES
  • During isolation procedure, tubes should be kept on ice or at 4°C
    • A refrigerated microcentrifuge is recommended
    • Use filter-tip pipettes that are sterile
  • After isolation, extracted RNA should be kept on ice or stored at -80°C
  • Never allow RNA to thaw on benchtop, always on ice!