BIO 2210 Introductory Biochemistry

Laboratory exercise 6
Date: March 14, 2017
Extracting DNA from green split peas

1 INTRODUCTION

The most recently discovered techniques in biotechnology are being conducted at the molecular
level. In order for you, the student, to gain an appreciation for this area, you may benefit from
actually seeing the genetic material which is manipulated through todays most state-of-the-art
techniques. For this activity, you will extract DNA from the cells of green split peas and cause it
to precipitate so that you can see the DNA with the naked eye.

2 MATERIALS & METHOD

Put in a blender: cup of green split peas; 1/8 teaspoon sodium chloride (NaCl); twice as much
cold water as split peas. Blend on high for 15 seconds to produce a thin pea-cell soup (blending
separates the pea cells from each other). Pour the thin pea-cell soup through a funnel lined into a
beaker. Measure the volume of pea soup in the beaker, and to this volume add about 1/6 of that
amount of liquid detergent and swirl to mix. Let the mixture sit for 10 minutes. Then, pour the
mixture into test tubes, each about 1/3 full. Add 3 drops of enzyme (contact lens solution) to each
test tube and gently stir the mixture using a stirring rod. Be careful, as stirring hard will break the
DNA molecules, making it harder to see. Following, tilt your test tubes and slowly pour alcohol
down the side of each tube, so that it forms a layer on the top of the pea mixture (pour until you
have about the same amount of alcohol in the test tube as the pea mixture). DNA will rise into the
alcohol layer from the pea mixture. Use a wooden stick, or hook to draw the DNA into the alcohol
(DNA is the long, stringy molecule that clump together). Finally, add 5 drops of phenol red/UI
indicator to the DNA solution and observe.

3 POST- LAB QUESTIONS

These questions MUST be submitted as a substitute of your Discussion.

1. Describe in your own words, what the extracted peas DNA look like.
2. What does the colour change with the phenol red/UI indicator tell you about the DNA
molecule?
3. DNA dictates the form and function of an organism. How does it do this?
4. Recombinant DNA technology is a modern day biotechnology application that allows
scientists to combine specific DNA sequences (gene) from one organism with the DNA
sequence of another organism.
a. In your own words and from your research, give logical reasoning as to why a
scientist might want to add DNA from one organism to another?
b. If you could use recombinant DNA technology to change the DNA of green split
peas, what new characteristics would you want it to have? Why?
c. Would you insert a DNA sequence from another plant or animal to create this new
pea? Explain.

BIO 2210 Kalicharan, L. (2017)

 5. Other biotechnology applications (similar to recombinant DNA) allow scientists to turn-
off or remove a DNA sequence.
a. Why might a scientist want to remove a DNA sequence from an organism?
b. If you could use an application of biotechnology to remove a specific characteristics
of green split peas, what characteristics would you want to remove? Why?

4 OUTPUT

A printed INDIVIDUAL report to be submitted on Tuesday, March 21, 2017, upon entering your
lab session. Your report MUST be typed in font: TNR, size: 12, justify, line spacing 1.5 on letter
size page (2.54 cm margins).

List of headings to be incorporated in your report:

1 INTRODUCTION No more than 1 pages based on nucleic acids and biotechnology and
the experiment conducted. Please be reminded that your AIM/OBJETCIVES are
subheadings of the introduction.
2 MATERIALS & METHODS Methodology must be written in a paragraph form and in the
past tense.
3 RESULTS: A short paragraph of your observations at each step.
4 POST-LAB QUESTIONS
5 CONCLUSION
6 REFERENCES Must be done in the APA Style.

Please DO NOT start each heading on a new page and if possible, kindly print back and front, if
you can.

BIO 2210 Kalicharan, L. (2017)