RNA Extraction and Handling

DOs

DON’Ts

  • Designate a special area for working with RNA
  • Clean benches with special cleaning solutions e.g. RNaseZap
  • Wear gloves and change them frequently
  • Use sterile, disposable plasticware
  • Glassware should be treated with DiEthylPyroCarbonate (DEPC) and oven baked at 180°C for at least 4 hours before use
  • Use filtered pipette tips
  • Solutions (water or other solutions) should be prepared with 0.1% DEPC
  • Use RNase-free reagents, tubes and tips
  • For best results use either fresh samples or samples that have been snap frozen on dry ice or in liquid nitrogen; all required reagents should be kept on ice
  • Store prepared RNA at -80°C
  • To avoid condensation of moisture during thawing, the slides should be frozen at -80°C and rethawed in a tightly sealed container (e.g. 50 ml Falcon tube)
  • Don’t breathe on samples; some researchers wear masks
  • Don’t touch anything with bare hands
  • Don’t autoclave pipette tips, as water vapor may contain RNases
  • Don’t allow frozen tissue to thaw
  • Don’t resuspend RNA in DEPC water; residual DEPC can inhibit downstream reaction
  • Don’t thaw and freeze frequently