DNA EXTRACTION

ontent Standard C: Life Science 9-12

• The Cell. Cells store and use information to guide their functions. The genetic information stored

in DNA is used to direct the synthesis of the thousands of proteins that each cell requires.

• The Molecular Basis of Heredity. In all organisms, the instructions for specifying the

characteristics of the organism are carried in DNA, a large polymer formed from subunits of four

kinds (A, G, C, and T). The chemical and structural properties of DNA explain how the genetic

information that underlies heredity is both encoded in genes (as a string of molecular “letters”) and

replicated (by a templating mechanism). Each DNA molecule in a cell forms a single

chromosome.

WHYS IT COLD

The cold ethanol helps the DNA to precipitate out of solution.

Strawberries have large genomes; they are octoploid,

which means they have eight copies of each type of chromosome in each cell.

The function of the DNA extraction buffer ingredients are as follows: (1) The soap helps to

dissolve the phospholipid bilayers of the cell membrane and organelles, (2) the salt is used to

break up protein chains that bind around the nucleic acids, and (3) the ethanol is used to

precipitate the DNA. DNA is not soluble in ethanol. DNA is least soluble in cold ethanol, so

make sure to keep the ethanol in the freezer or on ice. When the students add ethanol to their

strawberry extract, they will see the fine white strands of DNA precipitate. The DNA will form

cotton like fibers that will spool onto the stirring rod/inoculating loop/popsicle stick. Note:

More DNA will be obtained with ethanol than isopropyl alcohol.

How do we isolate DNA from the cell?

2. Ask students where DNA is found.

3. Ask them what the four main steps were of the DNA extraction.
• Crushed the cells to release the DNA. ee

• Used the buffer to separate the DNA from the other cell components.

• Filtered out the large particles.

• Precipitated out the DNA using ethanol.

4. Ask the students what the DNA looked like.

WHY STRAWBERRIES

Strawberries are a good source of DNA because they have 8 copies of each type of chromosome. This
large number of chromosomes will filter out of your solution and you will get to actually see DNA.

Review:

Where in the cell is the DNA found?

Take a look at the sketch of the plant cell below. The chromosomes (which are made of DNA)

are in the nucleus. This is the place where most of the DNA is located. (There are also small

amounts of DNA in the chloroplast and mitochondria.

Part I: Questions

a. What was the purpose of mashing up the strawberry?

To break down the cell wall, cellular and nuclear membranes.

b. What does the extraction buffer do? (Hint: Extraction buffer contains soap.

What does soap do when you wash your hands?)

The extraction buffer helps to release the DNA from the surrounding cell components

of the crushed [Link] soap helps to dissolve the phospholipid bilayers of the cell membrane and
organelles.

c. What does the filter do?

The filter removes the larger particles from the solution, such as seeds, pith, etc.,

allowing only the smaller cell components such as the DNA, proteins, etc. to filter

through.

Page 7:

Questions Parts II and III:

c. What happened when you added the filtrate to the alcohol?

The DNA precipitated out of the solution.

d. What did the DNA look like?

The DNA looked like white, thin fibers wadded up together forming a clump.

Pages 8-9:

1. What does DNA look like?

The DNA will look like a white, cloudy or fine stringy substance.

2. A person cannot see a single cotton thread four classrooms away. But if you

wound thousands of threads together into a rope, it would be visible at the same

distance. How is this statement an analogy to our DNA extraction?

DNA is not visible as a single strand to the naked eye, but when thousands of

threads of DNA are present, you will be able to see the large groups of threads of

3. Is DNA found in all living or once living cells?

Yes, DNA is present in all living and once living cells. Scientists can use DNA

from mummies or seeds that are thousands of years old and identify the genetics

of those plants and animals using their DNA.

4. Since the strawberries were once living, and we extracted DNA from them, what

does this mean about the foods you eat?

All the foods we eat that come from plants and animals that contain DNA. DNA

has been consumed on a daily basis since the beginning of time and is completely

safe to eat.

5. Look at the plant cell pictured on page one of the lab handout. Remember that

genes are found on chromosomes, and genes control traits. Give at least two

examples of traits that are expressed in the strawberry.

Examples: Color, size and sweetness.

6. Since we are studying about Bt and non-Bt corn in this unit, name at least two

traits a scientist would be interested in having in corn.

Examples: Yield, drought resistance, insect resistance, disease resistance, stalk

strength, root strength, early growth, stays green, days until maturity, and height.

(c) Separating the nucleic acids from the rest of the cells.

8. Use the filter paper/blotting paper/paper towel/(white)cloth/tea strainer (very fine sieve) and

funnel to filter the mixture, Larger particles/residues of the onion cells, such as cell walls

are left behind in the filter paper/blotting paper/paper towel/(white)cloth/tea strainer (very

fine sieve). Small particles including DNA and proteins pass through the filter paper

into the small beaker/bottle.

Unwrapping the nucleic acids from its histone proteins.

 1. Identify the DNA you extracted whether is most likely to be nuclear DNA, mitochondrial DNA
or chloro-plastic DNA. Give THREE reasons for your answer.
2. What precautions must you take in carrying out stages (d) and (e)?
3. State TWO substances that were used with water to prepare a solution.
4. Why is the experiment placed in the container with ice-cubes?
5. . Suggest TWO reasons why extreme cold methylated spirit is used.
6. State FOUR genetic materials used to detect culprits/criminals through DNA
7. 9. If the DNA molecule extracted was analysed and found to contain 14% of Thymine(T). What
percentage of the molecule is Cytosine (C)?
8. 10. State TWO functions of the DNA molecule?

Why do we add Salt?

Salt is used to break up protein chains that bind around the nucleic acids.

B Dishwashing liquid?

nd dish

soap to dissolve the lipid (fat) part of the strawberry cell wall and nuclear membrane. This

extraction buffer will help provide us access to the DNA inside the cells.

2.3 Give ONE reason why the ethanol should be kept on ice.

The colder the ethanol, the less soluble the DNA will be in it. Thus make sure to keep the ethanol in
the freezer or on ice.

(1)

2.4 State a conclusion to your experiment.

(2)

2.5 Draw a diagram of your results

Anna wants to identify her biological father. Three men were identified as being her possible father. A
DNA sample from her, each of the men and her mother were analysed. The diagram below was
produced:
4.1 Name the technique that was used to identify the father. (1)

4.2 Who is the possible father? (1)

4.3 State TWO other uses of the technique identified in QUESTION 4.1. (2)

PROCEDURE

A. Filter strawberry slurry through cheesecloth

B. Mush strawberry with salty/soapy solution

C. Initial smashing and grinding of strawberry

D. Addition of ethanol to filtered extract

FUNCTION

___ To precipitate DNA from solution

___ Separate components of the cell

___ Break open the cells

___ Break up proteins and dissolve cell membrane

5. Why is it important for scientists to be able to remove DNA from an organism? List two

reasons.
6. Is there DNA in your food? ________ How do you know?

Safety:

Ingestion of or contact with ethanol (or isopropyl alcohol) should be avoided. If contact with

the eye occurs, flush immediately with water. Inhalation of fumes or dermal contact with

ethanol (or isopropyl alcohol) is of minimal concern.

Erwin Chargaff a biochemist, researched the nitrogenous bases in DNA. His research

revealed the percentage of each base found in an organism’s DNA. The table below

includes some of his data.

1.1 What conclusion can be made about nitrogenous bases from the table above? (2)

1.2 Determine X. Show your workings.

The diagram below represents a nucleus and cytoplasm where several metabolic processes take place:
2.1 Identify processes:

A ______________________________________ (1)

B ______________________________________ (1)

2.2 Tabulate TWO differences between the two processes mentioned in

QUESTION 2.1.

Use the following shapes to draw a nucleotide present in mRNA, but not in DNA.
The Amino acid chart below is used to determine the specific amino acid by matching the

mRNA codons.

To find the amino acid encoded by the codon AGU:

1. Look up A in the 1st base (left side). This gives you the third block of amino acids

from the top.

2. Look up G in the 2nd base (top row). This gives you the fourth column of the third

block of amino acids from the top (Ser, Ser, Arg)

3. Look up U in the 3rd base (right side). This gives you the top amino acid in the

fourth column of the third block of amino acids from the top, i.e. Ser (serin
The first six nucleotides of the haemoglobin gene in DNA are:

CAC GTA

3.1 Determine the nucleotide sequence of the mRNA. (1)

_______________________________________________________________

3.2 Use your answer in QUESTION 3.1 to name the TWO amino acids that will be

translated from the m-RNA.

(2)

_____________________________ and ______________________________

3.3 Suppose the DNA sequence CAC GTA was changed to CAC GGA.

How would the products of transcription and translation be affected?

3.3.1 mRNA (2)

_________________________________________________________

3.3.2 Amino acid sequence (2)

 _________________________________________________________

3.4 Use the Amino Acid Chart to determine the TWO anticodons for the amino acid

Histidine.

(2)

_______________________________________________________________