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!