RNA extraction

RNA extraction

Protocol for 1 ml Trizol sample

  1. Add 0.2 ml chloroform.
  2. Shake tube to mix.
  3. Incubate at RT for 3 min.
  4. Centrifuge at 14,000 rpm at 4C for 15 min.
  5. Move supernatant to new 1.5 ml tube.
  6. Add 0.5 ml isopropanol.
  7. Mix by vortex.
  8. Centrifuge at 14,000 rpm at 4C for 10 min.
  9. Remove supernatant by decantation.
  10. Add 1 ml of 70% EtOH.
  11. Centrifuge at 14,000 rpm at 4C for 5 min.
  12. Remove supernate by 200 ul pipet.
  13. Dry up.
  14. Dissolve in 10-50 ul H2O
  15. Use 1 ug RNA for cDNA synthesis.
  16. Store at -80C.