SEQUENCING REACTION CLEANUP FOR ABI
 
  1. Add the following to the sequencing reaction:

    2.  
    50 µl 0.4M sodium acetate pH 4.6-5.2
    50 µl chloroform (if mineral oil was used)
    Vortex and centrifuge at 13,000 x g for 3 minutes

     
  2. Remove the aqueous (upper) phase to a fresh tube and add 150 µl 95% ethanol. Vortex and place on ice for 10 minutes.
  3. Centrifuge the reactions in the cold room for 20 minutes at 13-15,000 x g in the microcentrifuge.
  4. Remove the supernatant; be careful not to disturb the pellet (you do not need to remove all traces of the supernatant). Add 50 µl 95% ethanol. Centrifuge at 13-15,000 x g at room temperature for 5 minutes. Remove the supernatant without disturbing the pellet.
  5. Dry the tubes on your bench for 5-10 minutes or under vacuum for 5 minutes.
Resuspend pellet in 3.5 µl of loading buffer, heat at 95ºC for 3 minutes and run on gel.


Updated 01 December 2000.