Biology Lab: Simple DNA Extraction

Introduction: All living cells contain DNA. DNA is composed of long chains of genes. Genes contain the
protein building instructions for the cell. It was first determined in the 1940s that DNA was the carrier of the genetic information, but it was not until 1953 that Watson and Crick determined the structure of the DNA molecule. Today, much is known about how DNA codes and replicates information, and how that information is expressed in the phenotype of an organism. In this lab, we will attempt to remove the DNA from the cells of wheat germ. Wheat germ is composed of the embryonic cells of wheat seeds. Warm water will be used to soften the plant cell walls and ordinary dishwashing detergent is used to break up the membranes, allowing the DNA to precipitate in the solution.

Problem: What does DNA look like? Can DNA be extracted from the nuclei of eukaryotic cells? Materials Needed:
Wheat Germ 50 mL beaker Warm water bath Beral pipet Ice cold ethyl alcohol Balance Wood splint Graduated cylinder Dawn dishwashing detergent Microscope and microscope slides

Safety Precautions:
1. 2. Laboratory aprons and safety goggles are required for this lab. Always use caution when using laboratory chemicals. Never touch, taste or smell chemicals unless instructed to do so.

Procedure:
1. 2. 3. Use the balance to weigh out approximately one gram of wheat germ. Place the wheat germ in a small 50 mL beaker. Locate the water bath. This water is being kept at a temperature between 50 60 C. With a small beaker, dip a small amount of the water out of the water bath. Measure 20 mL of the warm water with a graduated cylinder. Pour the warm water into your test tube or beaker containing the wheat germ. 4. Using a wooden splint or glass stirring rod, mix constantly, but gently, for three minutes. 5. Use a graduated Beral pipet to obtain 1 mL of Dawn dishwashing detergent. Release the detergent into the beaker with the wheat germ. 6. Stir gently for 5 minutes, being careful not to create any foam. (If you do create foam, use a dropper or pipet to remove the foam.) 7. Use a graduated cylinder to measure 15 mL of ice-cold ethyl alcohol. 8. Hold the beaker of wheat germ at a 45-degree angle, and very slowly pour the alcohol down the side of the beaker. If poured correctly, the alcohol will form a layer on top of the wheat germ and detergent mixture. Do not mix the layers together. The DNA will precipitate at the boundary between the alcohol and the water. If the water layer and the alcohol layer mix together, the DNA will not precipitate. 9. Do not move the beaker for at least 15 minutes. White, stringy filaments of DNA will begin to appear at the water / alcohol boundary. If you let the solution sit undisturbed for 15 minutes, the DNA will begin to float to the top. 10. Use the wood splint or glass stirring rod to begin to spool the DNA. Very gently swirl the wood splint through the DNA. You should be able to wind the DNA around the wood splint. 11. To get more DNA to precipitate, dip the wood splint very gently down into the water layer, lifting the water up into the alcohol layer. 12. Place a small amount of your DNA on a clean microscope slide. Add a drop of methylene blue and a coverslip. Observe under low and high power.

Observation Questions:
1. What purpose is served by soaking the wheat germ in the warm water?

2.

What purpose is served by the Dawn dishwashing detergent?

3.

Describe what the DNA looks like with your naked eye.

4.

Describe what the DNA looks like under the microscope.

5.

Where did the DNA first appear?

6.

What might have happened if you poured the alcohol too fast and it did not form a layer on top of the water?

7.

There is about 6 feet of DNA contained in each of your cells. How is it possible for so much DNA to fit inside the small space of a cell?

8.

Why is DNA referred to as a double helix?

9.

What are the building blocks that compose DNA? What are the three components to these building blocks?

10. Draw a strand of DNA that has the following base sequence on the left hand side: ATTCGCTCG. Use the base paring rules to construct the right hand side of the molecule. Label the location of the sugars, the phosphates, and the nitrogen bases. Indicate where you would find a hydrogen bond.

11. DNA is able to make an exact copy of itself. What is this process called? Why is it so important that DNA be able to make an exact copy of itself?

12. What role does each of the following enzymes play in the replication of DNA? a) DNA polymerase

b) Helicase

Teacher Preparation:
1. This is a quick and dirty DNA extraction. There are many other lab procedures for this that can be used to extract a purer sample of DNA. I like this procedure for my standard Biology I students, because it is simple and straightforward. It can be done quickly, with minimal lab materials, and students can easily extract the DNA. The temperature of the water is very important. It must be kept between 50 60 C. Tools that can be used to spool the DNA: Paper clips, wood splints, bamboo skewers, glass stirring rods. It is essential that the alcohol be ice cold. I keep it in the freezer prior to the lab. During the lab, I keep it in an ice bucket. Have students try a variety of other cell materials and compare them. I have also used strawberries, onions and liver. I have tried different brands of dishwashing detergent, and Dawn always works the best. Ethyl alcohol seems to give me the best results, but I have used isopropyl alcohol and this also gives good results.

2. 3. 4. 5. 6. 7.

Answers to Questions:
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. Warm water is used to soften the plant cell walls. Dawn dishwashing detergent is used to break up the membranes, allowing the DNA to precipitate in the solution. Student answers will vary. Student answers will vary. The DNA will first appear at the boundary between the warm water/detergent solution and the cold alcohol. If the alcohol is added too fast, the water and alcohol will mix together. If the water layer and the alcohol layer mix together, the DNA will not precipitate. The DNA is tightly wound around proteins called histones. Histones are then supercoiled into nucleosomes. DNA is referred to as a double helix because it is two sided and twists around in a spiraling fashion. DNA is composed of monomers called nucleotides. A nucleotide is composed of three parts: a sugar molecule, a phosphate molecule, and one of four nitrogen bases.

11. When DNA makes an exact copy of itself, it is called replication. Replication occurs prior to mitosis and meiosis. An exact copy of all chromosomes must go into new daughter cells. 12. DNA polymerase catalyzes the elongation of new DNA by the addition of new nucleotides at the replication fork. Helicase is the enzyme that untwists the double helix of DNA at the replication forks, separating the two strands and making them available as template strands.