3. DNA Extraction - Logistics

Time
40 minutes

Grouping
individual

Materials
Each student needs:

• 1 fresh strawberry (frozen strawberries also work fine although they are not nearly as much fun to eat)
• 1 ziplock bag
• 1 15 ml centrifuge tube or 5 oz paper bathroom cup
• 1 clear glass or plastic test tube
• 1 paper towel
• 10 ml extraction buffer (see recipe below)
• 1 bamboo skewer or glass stirring rod (DNA tends to stick more fiercely to bamboo than wood – however, bamboo is MUCH cheaper)

Extraction buffer recipe:

• 450 ml distilled water
• 10 g table salt
• 50 ml Dawn dishwashing detergent

Optional for making necklaces:

• 1 1.5 ml Eppendorf tube (also known as microcentrifuge tubes, available from Science Kit and Boreal Labs at $13.25 for 500 tubes)
• 50 cm of string or yarn

For the whole class to share:

• clean food bowl
• clean toothpicks for eating strawberries
• 90% ice cold rubbing alcohol or ethanol
• eye droppers for dispensing solutions

Setting
Classroom

3. DNA Extraction - Background

Teacher Background
This activity should be part of the standard repertoire of any teacher who teaches genetics. It is essential for students to prove to themselves that DNA exists and that it can be extracted from any cell. Strawberries are used in this activity because they are octaploid, meaning they have 8 copies of every gene rather than the usual 2; thus providing prodigious quantities of DNA to extract. Naturally, strawberries are also relatively inexpensive and readily available. Other sources of DNA to experiment with include kiwis, bananas, and calf thymus.

The DNA molecule is an invisibly thin, very long strand. The DNA found in each human cell is almost 2 meters long. If all the DNA in a human adult (that’s 100 trillion cells) were laid end to end, the DNA would stretch 113 billion miles. That would take you to the sun and back 610 times. Even though DNA is invisible to the naked eye, no microscopes are needed! The reason is that you release so many DNA strands that they tangle together into a thick cable, visible without magnification. For example, it would be the same as if you took a thin piece of thread and held it up on the far end of the hallway. You probably wouldn’t be able to see the thread from that distance. However, if you took the thread and tangled it up with a hundred thousand other threads, you would be able to see the tangled clump from far away because there is so much of it.

The process itself is fairly straightforward. First the cell walls are broken open by smashing the strawberries in a ziplock bag. Next, detergent is used to dissolve the cell and nuclear membranes. The membranes are made of lipids (fat) and the detergent will cut through the membrane just like it cuts through grease on a dirty plate when washing dishes. Some salt is present in the detergent solution in order to match the osmolarity of the cells.

Now you have a big mixture of smashed cell walls, dissolved membranes, loose DNA and random other cell parts. This mixture is filtered through paper towels. Finally, you take advantage of the fact that DNA is soluble in water but not in alcohol. In fact, alcohol makes DNA clump together. Thus a layer of alcohol laid on top of the filtrate. Any DNA that contacts the alcohol will clump together, pulling the rest of the DNA strand along behind it. Soon you should see gossamer white strands of DNA bubbling their way up from the red strawberry extract.

The DNA may be collected by twirling a bamboo skewer or glass stirring rod in the solution. The DNA will spool itself around the skewer and can be pulled out of the solution. To keep some DNA, students may fill an eppendorf tube with alcohol and place their spooled DNA into the container. Lay the string on the hinge holding the cap to the tube and close the lid. The string forms a necklace with the eppendorf and enclosed DNA as a pendant. Top off the alcohol in the pendant and you can keep the DNA indefinitely.

Student Prerequisites
Some cell biology experience (enough to know that DNA is located in the nucleus of a cell and that membranes are made of lipids) is useful. If students are not aware of these fact, expect to spend at least 10 minutes longer teaching these ideas before starting the extraction.

3. DNA Extraction - Getting Ready

Getting Ready

1. Purchase strawberries, enough for each student to have one. (They will eat half and use the remainder to extract DNA.)
2. Prepare the extraction buffer.
3. Put the alcohol in the freezer or on ice.
4. Wash the strawberries and remove the green tops. Cut each strawberry in half. Put half in a clean food bowl for students to eat. Put the remainder in a separate bowl for students to extract DNA from.
5. Set out the remainder of the materials.

3. DNA Extraction - Lesson Plan

Lesson Plan

1. Have students write down a few sentences to describe what DNA is and what they think DNA looks like. After this lab or the series of DNA modeling activities, they will come back to this naive description to revise their answers with a more scientific one.
2. Draw a diagram on the board showing DNA (as a long tangled thread) within the nucleus of a cell. Label the DNA, nucleus, cell membrane, and cell wall. Remind (or teach) students about basic cell structure.
3. Tell students that they will be extracting the DNA from a strawberry and will then be able to look at the DNA. Briefly describe the process explaining the purpose of each of the steps.
4. Pass out ziplock bags and strawberries. Tell students there are strawberries to eat after the lab is cleaned up.
5. Students should put the strawberry in the bag, squeeze out most of the air and seal the bag. The strawberry can then be crushed into juice and pulp. Try to squish all of the chunks into an even, smooth puree. Warn students not to pound the strawberry on the table or risk the bag bursting and getting strawberry pulp all over themselves and the classroom.
6. Next, open the bag and add 10 ml of extraction buffer (approximately 10 eyedroppers full). Seal the bag again and gently mix the strawberry juice with the extraction buffer. Warn students not to mix too vigorously or it will generate a lot of bubbles and can’t be filtered effectively. Use a gentle tilting back and forth motion while lightly squeezing the bag.
7. Set up a filtration system. I had students wrap a paper towel around their finger then put their paper-wrapped finger into the mouth of the 15 ml tube or 5 oz cup. When you remove your finger, the paper towel should form a well into which the strawberry juice can be poured.
8. Carefully pour the extract into the well in the paper towel. Allow the juice to filter through the towel into the container below. Let it drip for 3-5 minutes. Do not squeeze the towel or you will create lots of bubbles, disrupting the interface needed in the next step.
9. The paper towels can be put inside the ziplock bags and thrown away.
10. Carefully transfer liquid from the 15 ml tube or cup into the clear test tube until the test tube is about a third full.
11. Slowly add 3 ml (3 eye droppers full) of ice cold alcohol to the test tube. The alcohol should be added so that it trickles down the side of the tube before pooling on top of the strawberry extract. You should end up with a red bottom layer and a clear top layer.
12. Have the students make observations of anything going on in the clear alcohol layer. You may wish to have students write down observations at this point.
13. After 2-3 minutes, a skewer or stirring rod can be inserted into the tube and gently swirled around. This will spool the DNA around the stick. The DNA can be pulled out of the tube and stored in a microcentrifuge tube filled with some alcohol. Students may safely touch the DNA although the DNA should NOT be tasted under any circumstances.
14. By trapping a piece of string in the lid of the microcentrifuge tube, students can wear their DNA home as a necklace.
15. Have students clean up their areas. Nicely cleaned tables and washed hands may be rewarded with a piece of strawberry to eat.

3. DNA Extraction - Assessment

Assessment

1. Have students answer summary questions about the extraction.

• Why is it necessary to mash the strawberries?
• What is the purpose of the detergent?
• What is the purpose of the salt?
• Name a liquid that DNA is not soluble in.
• Is the DNA that you extracted pure? What else might be attached to the DNA?
• Why might some people get more DNA than others?
• Can you see a single strand of DNA without a microscope? Explain how you were able to see the DNA in this experiment without magnification.

Going Further

1. Make models of DNA (see DNA Models lesson).
2. Try to isolate DNA from other soft fruits and vegetables. This may even be done as homework. Compare the DNA yields and discuss why different plants would give different results.

3. DNA Extraction - Sources and Satndards

Sources
There are numerous write ups for this experiment available on the internet and elsewhere. I first experienced this lesson through UCSF’s Science and Health Education Partnership. I then tried it with Carolina Biological .

The estimates of the length of DNA in a human cell and the number of cells in the human body were taken from Wikipedia (from the Genome and Cell Biology articles).

Standards
Grade 7
Genetics
2. A typical cell of any organism contains genetic instructions that specify its traits. Those traits may be modified by environmental influences. As a basis for understanding this concept:
e.    Students know DNA (deoxyribonucleic acid) is the genetic material of living organisms and is located in the chromosomes of each cell.

Grades 9-12
Genetics
5. The genetic composition of cells can be altered by incorporation of exogenous DNA into the cells. As a basis for understanding this concept:
a. Students know the general structures and functions of DNA, RNA, and protein.