DNA Extraction from Bacteria

Materials:

E. Coli suspension (Luria Broth, 37^oC overnight)
1% Sodium Dodecyl Sulfate (SDS)
Test tubes
Water Bath (60-65^oC)
Stirring rod
95% Ethanol (chilled)
Dropper

Safety: Be careful working with Bacterial culture ! Wash hands!

Procedure:

1. Add 5 ml. E. Coli suspension to a test tube.

2. Add 1 ml. of SDS to the Tube with E. Coli, gently rotate and swirl to mix in SDS approx. 5 min.

3. Stand tube in a 60-65^o C water bath for 30 min.

4. Allow to cool to room temp.

5. Place a stirring rod into the test tube and add 2 ml. of cold 95% ethanol slowly with the dropper down the stirring rod into the tube.

6. Hold tube at a 45 degree angle and slowly rotate your stirring rod clockwise (avoid touching sides of tube) approx. 5 min.

7. Remove rod and immerse rod with bacterial DNA in 95% ethanol for approx. 2 min.