Thursday, March 7, 2013

DNA Extraction from Wheat Germ

Instructions:

1. Place 1 gram of raw wheat germ in 50 ml test tub.

2. Add 20 ml of hot (50-60 C) tap water and mix constantly for 3 minutes.

3. Add 1 ml or a scant 1/4 teaspoon of detergent and mix gently every minut for 5 minutes. Try not to create foam.

4. Use an eyedropper, pipette, or piece of paper towel to remove any foam from the top of the solution.

5. Tilt the test tube at an angle. SLOWLY pour 14 ml of alcohol down the side so that it forms a layer on top of the water/wheat germ/detergent solution. Do not mix the two layers together. DNA precipitates at the water-alcohol interface (the boundary between the water and the alcohol). Therefore, it is crucial to pour the alcohol very slowly so that it forms a layer on top of the water solution. If the alcohol mixes with the water, it will become too dilute and the DNA will no precipitate.

6. Let the test tube sit for a few minutes. White, stringly, filmy DNA will begin to appear where the water and alcohol meet. You will usually see DNA precipitating from the solution at the water-alcohol interface as soon as you pour in the alcohol. If you let the preparation sit for 15 minutes or so, the DNA will float to the top of the acohol.
     You can usually get more DNA to precipitate from the solution by using one of the DNA-collecting tools (such as glass or paper clip hook) to gently lift the water solution up into the alcohol. This allows more DNA to come in contact with the alcohol and precipitate. You may find it helpful to pour the water/detergent solution into a clean test tube, leaving behind the wheat germ, before adding the alcohol.

7. Use a glass or paper clip hook or wooden stick to collect the DNA.

OBSERVATIONS

1. What does the "wheat germ soup" look like?
     The "wheat germ soup" looks much like watery snot...interesting....different.

2. How does its appearance change as you add the detergent and swirl it in?
     The "snot look" begins to form. The DNA, or "stringy parts" start to develop.

3. What do you think is happening at this step?
     The DNA is beginning to form.

4. Describe the appearance of the mixture just after you add the alcohol.
     The two solutions are separate, but in the same tube. It looks layered.

5. What do you think is happening at this step?
     The two solutions are separating so that DNA can form in between them. They're working as glues.

6. What do you observe at the water-alcohol interface?
     The wheat germ met the alcohol, so that's where the filmy DNA film started forming.

7. What does DNA look like?
     The DNA looks like snot. Creation of life is beautiful...but really not too pretty!