Developed by Darla French



An Apple a Day:

Extracting DNA from Any Living Thing

EXTENDED VERSION

Unit: The Apple Genomics Project

Lesson: An Apple a Day: Extracting DNA from Any Living Thing

Audience: This lesson is intended for use with middle and high school science students.

Student Learning Objectives:

At the completion of this lesson the students will be able to:

1. Define the process of DNA extraction.

2. Explain the purpose of the extraction process.

3. Explain why DNA is essential and a part of all living things.

4. Explain where DNA is found.

5. Demonstrate the ability to follow instructions in a lab experiment.

6. Explain the function of each material in a lab experiment.

7. Compare results of the exercise using different sources for DNA extraction.

Student Activities:

• Take notes on lesson/pre-lab content as presented by instructor.

• Complete pre-laboratory worksheet.

• Complete laboratory exercise.

• Discuss laboratory exercise.

• Complete post-laboratory worksheet.

Problems & Questions for Study:

1. Where is DNA found?

2. Why is DNA essential to all living things?

3. What is DNA extraction?

4. What is the purpose of the extraction process?

5. What are the steps in the DNA extraction process?

6. What is the function of each material in this lab experiment?

7. How do the results of the exercise change, using different sources for DNA extraction?

Documentation of Competencies and Academic Standards Met:

National Academic Standards

Science Content Standard A for Grades 9-12: Develop abilities necessary to do scientific inquiry.

Science Content Standard B for Grades 9-12: Develop understanding of chemical reactions.

Science Content Standard C for Grades 9-12: Develop understanding of the cell.

Connection to Supervised Agricultural Experience/Career Development

Careers related to horticulture include fruit grower, extension agent, plant cytologist, flower grader, bacteriologist, grounds keeper, and tree surgeon. [For more careers, visit the FFA Career Center at .]

FFA proficiency areas related to horticulture include floriculture, landscape architecture, and specialty crop production. [For more proficiency areas, visit the National FFA website for proficiencies at .]

Connection to FFA/Leadership Development/Personal Growth

Career Development Events (CDEs) related to horticulture include floriculture and nursery & landscaping. [For more information on CDEs, visit .]

Motivation/Interest Approach:

Display various items to the class, including “living”, “never living”, and “previously living” objects. Ask students to explain what DNA is. Once the class comes up with a definition of DNA, ask students to identify which objects contain DNA. Have students sort the objects into the three categories (living, never living, or previously living) based on their identifications. Ask students to explain how and why they categorized each object the way they did. Most likely, students will sort objects into only two categories: “living” and “non-living”. If this is the case, point out that some of the objects were “previously living”. Ask students whether these objects have DNA, since they were alive at a previous time. Initiate a discussion in which students may re-categorize objects, if necessary, to include “living”, “never living”, and “previously living” objects. Ask students how they know that “living” cells (and “previously living” cells) contain DNA, if they cannot see physical proof of the DNA itself. Explain to students that the laboratory exercise in this lesson will allow them to extract and observe DNA from living cells.

Content Outline & Teaching Procedures:

I. Background

A. DNA ~ deoxyribonucleic acid

B. DNA is present in EVERY cell of ALL living organisms.

C. Long, thick fibers, arranged in a double helix, of DNA store information for the functioning of the chemistry of life, and for building and maintaining a living organism.

D. The process of extracting (taking it out of, or removing it from) DNA from a cell is the first step for many laboratory procedures in biotechnology, including gel electrophoresis and DNA sequencing.

1. Scientists use DNA extraction to separate DNA from the unwanted substances of the cell, in order to examine the DNA.

2. It must be done gently enough so that the DNA does not denature (break up).

II. Laboratory Exercise

A. DNA is the “blueprint for life,” and it can be extracted from any living (or formerly living) thing.

1. Examples of things with DNA that we could extract: Split peas, spinach, chicken liver, onions, and broccoli. (We’ll use bananas for this exercise, because they have a LOT of DNA packed into cells that are easy to break open.) All of these are things that are living.

2. Examples of things with DNA that we could extract, but probably would not want to try extracting: Wood, preserved fossils, cotton. These are things that are living, or were previously living, but DNA extraction would be difficult.

3. Examples of things with no DNA: Rocks, water, candles, plastic, and pop cans. All of these are things that never lived.

B. DNA found in banana cells can be extracted using common materials. This procedure uses household equipment and store supplies to extract DNA from bananas in sufficient quantity to be observed and spooled.

1. In this exercise, you will prepare a solution of banana treated with salt, distilled or tap water, and Dawn dishwashing detergent (or Suave clarifying shampoo).

a. Salt breaks up the protein chains that bind the nucleic acids. The salt allows the DNA to precipitate out of a cold alcohol solution.

b. Dawn dish soap is a detergent, so it dissolves the lipid (fat) part of the banana cell wall and nuclear membrane, allowing the DNA inside to spill out.

i. The detergent breaks down the cell membrane by dissolving the lipids (fatty molecules) and proteins of the cell and disrupts the bonds that hold the cell membrane together.

ii. The detergent then forms complexes with these lipids and proteins, which are filtered out of solution by the coffee filter, while leaving the cells' DNA in the filtrate (the liquid solution that goes through the coffee filter into the cup).

c. A blender will be used to separate the banana cells from each other, resulting in a thin banana soup.

2. Successful spooling will result in clumps of white, stringy DNA.

a. The white, stringy stuff is actually a mix of DNA and RNA.

b. The procedure for DNA extraction is really a procedure for nucleic acid extraction. Much of the RNA is cut by ribonucleases (enzymes that cut RNA) that are released when the cells are broken open.

C. Troubleshooting the laboratory exercise

1. Problem: No DNA precipitates out of banana/detergent/alcohol solution.

a. Solution 1: First, check one more time for DNA. Look very closely at the alcohol layer for tiny bubbles; clumps of DNA are often loosely attached to the bubbles.

b. Solution 2: If you still do not see DNA, then you need to make sure that you started with enough DNA in the first place. Many food sources of DNA, such as grapes, also contain a lot of water. If the blended cell soup is too watery, there will not be enough concentrated DNA to see. To fix this, go back to the first step and add less water. The cell soup should be opaque (you should not be able to see through it).

c. Solution 3: If you still do not see DNA, then you need to make sure that you allow enough time to pass for each step. Make sure to stir in the detergent for at least five minutes. If the cell and nuclear membranes are still intact, the DNA will be stuck in the bottom layer. Often, if you let the banana/detergent/alcohol mixture sit for 30-60 minutes, DNA will precipitate into the alcohol layer.

2. Problem: DNA clumps together.

a. Solution: None. Single molecules of DNA are long and stringy. For example, each cell of your body contains six feet of DNA, but it's only one-millionth of an inch wide. To fit all of this DNA into your cells, it needs to be packed efficiently, so DNA twists tightly and clumps together inside cells. Even when you extract DNA from cells, it still clumps together, though not as much as it would inside the cell.

D. Using this DNA as a sample for gel electrophoresis.

1. You can try this, but all you will most likely see is a smear.

2. The DNA you have extracted is genomic, meaning that you have the entire collection of DNA from each cell. Unless you cut the DNA with restriction enzymes, it is too long and stringy to move through the pores of the gel.

E. Other things to have students try:

1. Experiment with other DNA sources. Which source gives you the most DNA? Why do you think this is the case? How can you compare them?

2. Experiment with different soaps and detergents. Try powdered soaps, clear shampoo, body scrub, meat tenderizer, pineapple juice, or contact lens cleaning

solution.

3. Try leaving out a step or changing how much of one ingredient you use.

References & Teaching Aides:

Websites:

• DNA in My Food? The Making of a Smoothie:

• The Science Behind Our Food: Lesson in a Trunk: Banana DNA Extraction:

• Genetic Science Learning Center at the Eccles Institute of Human Genetics at the University of Utah:

Books:

• DNA Is Here to Stay, by Dr. Fran Balkwill. 1992. ISBN # 0-00-191165-1. (Ages 9-15)

• Amazing Schemes within Your Genes, by Dr. Fran Balkwill. 1994. ISBN # 0876146353. (Ages 9-12)

• Double-Talking Helix Blues, by Joel Herskowitz. 1993. Book and cassette tape. ISBN # 0-87969-431-9. (Ages 8 and up)

• Ingenious Genes, by Patrick A. Baeuerle and Norbert Landa. 1998. Barron's Educational Series, Inc., Hauppauge, NY. ISBN # 0-7641-5063-4. (Ages 9-12)

• How the Y Makes the Guy, by Norbert Landa and Patrick A. Baeuerle. 1998. Barron's Educational Series, Inc., Hauppauge, NY. ISBN # 0-7641-5064-2. (Ages 8-12)

Additional Materials:

Handouts

An Apple a Day: Class Notes

Worksheets

An Apple a Day: Pre-Laboratory Worksheet

An Apple a Day: Post-Laboratory Worksheet

Laboratory Worksheets

An Apple a Day: Laboratory Exercise

An Apple a Day: Laboratory Exercise, Part 2

Name: ___________________________________

An Apple a Day: Class Notes

I. Background Information

A. DNA is the acronym for ______________________________ ________________.

B. DNA is present in _________________ cell of ________________ living organisms.

C. Long, thick fibers of DNA store _________________ for _______________________

______________________________________________, and for __________________

_______________________________________________________________________.

D. The process of extracting DNA from a cell is the first step for ___________________

_________________________________, __________________________________ and

__________________________________.

1. Scientists separate _________________ from the _______________________

________________________________________________________, in order to

___________________________________________________.

2. This must be done gently enough so that _______________________________

_________________________________________________________________.

II. Information about the Laboratory Exercise

A. Since DNA is the “__________________________________,” it can be extracted

from __________________________________.

1. Examples of things with DNA that we could extract: _____________________

_________________________________________________________________.

(We’ll use _________________ for this exercise, because they have a LOT of

DNA packed into cells that are easy to break open.) All of these are things that are

_________________.

2. Examples of things with DNA that we could extract, but probably would not

want to try extracting: _______________________________________________

__________________________________________________. All of these are

things that are _________________, or were _____________________________.

3. Examples of things with no DNA: ____________________________________

________________________________________. All of these are things that

__________________________________.

B. The DNA found in banana cells can be extracted using _________________________

______________________.

This procedure uses _______________________________________________________

and __________________________________ to extract DNA from bananas in sufficient

quantity to be _________________ and _________________.

1. In this exercise, you will prepare a solution of _________________ treated

with _________________, _________________, and _________________.

a. Purpose of salt: _____________________________________________

____________________________________________________________

___________________________________________________________.

b. Dawn dish soap is a type of _________________. Purpose of Dawn

dish soap: ___________________________________________________

____________________________________________________________

___________________________________________________________.

i. The detergent ______________________________________ by

_____________________________________________ of the cell

and __________________________________ that hold the cell membrane together.

ii. The detergent then forms ___________________________ with

_____________________________________, allowing them to be

________________________________________________ by the

_________________, while leaving the cells' _____________ in the filtrate.

c. We’ll use a __________________ to mix the banana solution. This step

will ______________________________________________, resulting in

___________________________________________________________.

2. Successful spooling will result in ____________________________________.

a. The white, stringy stuff is actually a mixture of _______ and _______.

b. The procedure for DNA extraction is really a procedure for __________

_____________________________________. Much of the RNA is cut by

____________________ (enzymes that cut RNA) that are released when

the cells are ____________________.

B. Troubleshooting the laboratory exercise

1. Problem: No DNA precipitates out of banana/detergent/alcohol solution.

a. Solution 1: ________________________________________________

____________________________________________________________

____________________________________________________________

___________________________________________________________.

b. Solution 2: ________________________________________________

____________________________________________________________

____________________________________________________________

___________________________________________________________.

c. Solution 3: ________________________________________________

____________________________________________________________

____________________________________________________________

___________________________________________________________.

2. Problem: DNA clumps together.

a. Solution: ________________________________________________

____________________________________________________________

____________________________________________________________

___________________________________________________________.

Name: ___________________________________

An Apple a Day: Pre-Laboratory Worksheet

1. Where is DNA found? _________________________________________________________

2. What do you think the extracted DNA will look like? ________________________________

______________________________________________________________________________

3. How could DNA that is extracted from a human be used? Name 3 ways:

a. ____________________________________________________________________________

b. ____________________________________________________________________________

c. ____________________________________________________________________________

4. The human body contains about 100 trillion cells. Each of these cells contains approximately six feet of DNA. To about how many total miles of DNA is this equivalent? Recall that 100 trillion = 1 x 1014, and 1 mile = 5,280 feet. Show your work below.

5. Why is salt used in this exercise? ________________________________________________

______________________________________________________________________________

6. Why is the Dawn dish soap (or Suave clarifying shampoo) used in this exercise?

______________________________________________________________________________

7. Why is alcohol used in this exercise? _____________________________________________

______________________________________________________________________________

8. What organism will provide DNA for extraction in this exercise? ______________________

9. A coffee filter will be used in this exercise. What do you think it will be used for? __________

______________________________________________________________________________

10. Circle the item(s) in this list that contain DNA:

Zebra fish Shale Ruby

Gum wrapper Pine tree Blood

Mosquito Wood Yogurt

Carrot Orange Cat

Quartz Candle Compact disc

Name: ___________________________________

An Apple a Day: Post-Laboratory Worksheet

Results, Analysis, and Conclusions

1. Describe the appearance of the suspension in the test tube. ____________________________

______________________________________________________________________________

Using the diagram below, sketch what the suspension in your test tube looked like after you completed the last step of the exercise. Be sure to label any layers of liquid there may have been, as well as where the DNA was:

2. Why was the salt used in the beginning step? _______________________________________

______________________________________________________________________________

______________________________________________________________________________

3. When you do dishes at home, what happens when you put dish soap in a bowl or pan with a

lot of grease (fat) in it? ___________________________________________________________

______________________________________________________________________________

4. Why was the detergent added to the banana mixture? _________________________________

______________________________________________________________________________

5. Why do you think it might be important for scientists to be able to remove the DNA from an

organism? _____________________________________________________________________

______________________________________________________________________________

6. Is there DNA in your food? ___________ How do you know? _________________________

______________________________________________________________________________

7. What do we mean when we say the DNA has been “extracted” from the banana cells? ______

______________________________________________________________________________

______________________________________________________________________________

8. List the steps of the DNA extraction process as you followed them in the exercise. What did the banana solution look like at each step?

STEPS OBSERVATIONS

9. How do you think the results of the exercise would change, if you had used a different

organism from which to extract the DNA? ___________________________________________

______________________________________________________________________________

______________________________________________________________________________

10. What is the purpose for extracting DNA? How might extracted DNA be used? ___________

______________________________________________________________________________

______________________________________________________________________________

______________________________________________________________________________

11. Where is DNA found? ________________________________________________________

______________________________________________________________________________

12. Why is DNA essential to all living things? ________________________________________

______________________________________________________________________________

13. Briefly explain the purpose of each of the following materials, as used in the exercise:

|Plastic cup | |

|Blender | |

|Plastic spoon | |

|Coffee filter | |

|Distilled water | |

|Detergent or shampoo | |

|Bananas | |

|Salt | |

|Plastic transfer pipette | |

|95% ethanol | |

|Small tube with cap | |

|Ice and ice bucket | |

Name: ___________________________________

An Apple a Day: Laboratory Exercise

In this exercise, you will extract DNA from bananas that have been blended with water. You will then treat a portion of the banana mixture with detergent, salt, and cold alcohol to precipitate the DNA from the banana solution.

Materials Needed

Blender

Two 5-ounce plastic cups

Plastic spoon

Graduated cylinder*

#2 cone coffee filters

Distilled or tap water

Dawn dish detergent**

2 peeled bananas

Teaspoon

2 pinches of table salt (iodized or non-iodized)

Plastic transfer pipette or medicine dropper

Two test tubes, with caps

Test tube stand

95% ethyl alcohol***

Ice and ice bucket

Glass rod****

Post-laboratory worksheet

*Or liquid measuring cup and dry measuring spoons

** Or Suave clarifying shampoo

*** Or 91% isopropyl alcohol

****Or wooden stick, or glass Pasteur pipette modified into a hook

Procedure

Carefully read and follow the directions. Place a checkmark next to each step as you complete it.

1. Obtain an ice bucket and fill it with ice.

2. Place about 5 mL ethanol (or isopropyl alcohol) into one test tube, cap the test tube, and place it (upright) into the ice bucket. You’ll need to use ice-cold alcohol to get the best results for this exercise.

3. Add two peeled bananas and one cup (about 250 mL) distilled water to a blender. Blend for 15-20 seconds, until the solution is a mixture. (The instructor may have already performed this step, so check first.)

4. In one of the 5 oz. plastic cups, make a solution of one teaspoon of Dawn dishwashing detergent and two pinches of table salt. Add 4 teaspoons (about 20 mL) of distilled water or until the cup is about one-third full. Dissolve the salt and shampoo by stirring slowly with the plastic spoon to avoid foaming.

5. Add three heaping teaspoons (about 15-20 mL) of the banana mixture from step three to the solution -from step 4. Slowly stir the solution with the plastic spoon for 5-10 minutes. (The detergent dissolves the lipids that hold the cell membranes together, thus releasing the DNA into solution. The detergent causes lipids and proteins to precipitate out of the solution, leaving the DNA. The salt enables the DNA strands to come together.)

6. Place a #2 coffee filter inside of the remaining 5 oz. plastic cup. Fold the coffee filter’s edge around the cup so that the filter does not touch the bottom of the cup.

7. Pour the mixture from step 5 into the filter. Let the solution drain for several minutes until there is approximately 5 ml (covers the bottom of the cup) of the filtrate, from which the DNA will be extracted.

8. Fill the plastic pipette with solution from step 7 and transfer it to the remaining test tube.

9. Remove the test tube of cold alcohol from the ice bucket. For best results, the alcohol should be as cold as possible.

10. Tilt the test tube containing the filtrate to one side. Slowly pour the alcohol into the filtrate down the side, so it forms a layer on top of the banana filtrate. Pour until you have about the same amount of alcohol in the tube as filtrate. Alcohol is less dense than water, so it will float on top of the banana solution.

11. Let the column sit for 2-3 minutes without disturbing it. It is important not to shake the tube during this time, because the strands of DNA are fragile at this point. DNA is not soluble in alcohol. When alcohol is added to the mixture, the components of the mixture, except for DNA, stay in solution while the DNA precipitates out into the alcohol layer.

12. You can watch the white DNA precipitate out of solution into the alcohol layer. DNA has the appearance of white, stringy mucus. The protein and grease parts from the banana cells stay in the bottom, watery layer of banana filtrate, while the DNA precipitates into the top layer of alcohol. Look for DNA at the point where the alcohol layer touches the filtrate layer, since this is where the DNA will precipitate first.

13. When good results are obtained, there will be enough DNA to spool on to a glass rod. Or, you can retrieve some of the DNA by using a wood stick or a Pasteur pipette that has been heated at the tip to form a hook. Once the DNA has risen into the alcohol layer from the banana layer, the DNA will be a long, stringy molecule that clumps together.

14. Congratulations! You have just extracted DNA! Now, answer the questions on your post-laboratory worksheet. Clean up your workspace according to your instructor’s directions.

Name: ___________________________________

An Apple a Day: Laboratory Exercise, Part 2

Make a Smoothie

With permission from your instructor, after you finish your extraction activity you may follow the recipe below to make a banana-strawberry smoothie:

1. To the remaining banana mixture in the blender, add the following ingredients and blend until smooth.

• An additional peeled banana, cut into pieces

• One 10-ounce package of frozen strawberries, partially thawed

• One 12.3-ounce package of tofu (either soft or firm, cut into pieces)

• One cup orange juice (add more for a thinner consistency)

• Two tablespoons of honey (or more, to taste)

2. Share the smoothie with your classmates in 5 oz. cups. This recipe makes enough for 10 5 oz. cups.

3. Evaluate your smoothie. Why do you think the tofu is in the smoothie?

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