Protocols

RNA extraction

  • 1. 1ml TRIzol per well. 5 min at RT.
  • 2. Add 200µl chloroform, shake 15 sec, incubate 3 min at RT.
  • 3. Cfg 15 min at ≤12000 g, 4C.
  • 4. Aqueous phase + 0.5ml isopropanol, incubate 10 min at RT.
  • 5. Cfg 10 min at ≤12000 g, 4C.
  • 6. Wash with 1 ml 75% EtOH, 5 min at ≤7500 g, 4C.
  • 7. Dissolve the pellet in µl DEPC H2O.
  • 8. Spec 1µl RNA + 99µl H2O (dilution 1:100).