How to isolate DNA – First Steps in

Biotechnology
Deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) are the two types of nucleic acids
found in living organisms. DNA acts as the genetic material in organisms. RNA, although it also
acts as a genetic material in some viruses, has different functions, perhaps the most important of
which is to ‘carry’ the genetic information from DNA to ribosomes, where proteins can be made.

OBJECTIVE
To isolate DNA from plant or animal tissue such as spinach leaves, green pea seeds,
strawberries, broccoli, papaya, chicken liver etc.

MATERIALS – collect everything together before you start

✔Plant or animal tissue (spinach leaves, green pea seeds, papaya, chicken liver, etc

✔Pestle & mortar ✔Beakers ✔Test tubes & TT rack

✔Enzymes (Meat tenderizer) ✔Cold ethanol ✔Salt ✔Sieve/strainer

✔Spooling stick ✔Liquidiser ✔Cold water ✔Liquid detergent

PROCEDURE

Step 1: Making a cell soup

1. Put the following in a blender:
• 1/2 cup of split peas or other plant or animal tissue (100ml)
• 1/8 teaspoon table salt (less than 1ml)
• 1 cup cold water (200ml)

2. Blend on high for 15 seconds. The blender breaks the tissue up and separates the cells from
each other, so you now have a really thin soup.

Step 2: Making a soapy, cell soup

1. Pour your thin soup through a strainer into another container (like a measuring cup).
2. Add 2 tablespoons liquid detergent (about 30ml) and swirl to mix.
3. Let the mixture sit for 5-10 minutes.
4. Pour the mixture into test tubes or other small glass containers, each about 1/3 full.
Step 3: Enzyme Power – breaking down the proteins which protect the DNA
Use meat tenderizer for enzymes. If you can't find tenderizer, try using pineapple or papaya flesh
juice or even contact lens cleaning solution. The enzymes you want are protein-digesting enzymes
(proteases), such as bromelain (from pineapple) or papain (papaya).
1. Add a pinch of enzymes to each test tube and stir gently. (Be careful! If you stir too hard, you'll
break up the DNA, making it harder to see.)

Step 4: Alcohol Separation

1. Tilt your test tube and slowly pour cold rubbing
alcohol (70-95% isopropyl or ethyl alcohol) into the
tube down the side so that it forms a layer on top
of the pea mixture. Pour until you have about the
same amount of alcohol in the tube as the cell
mixture. Alcohol is less dense than water, so it
floats on top. Look for clumps of white stringy stuff
where the water and alcohol layers meet.

What is that Stringy Stuff?

DNA is a long, stringy molecule. The salt that you added in step one
helps it stick together. So what you see are clumps of tangled DNA
molecules!

DNA normally stays dissolved in water, but when salty DNA comes in
contact with alcohol it becomes undissolved. This is called
precipitation. The physical force of the DNA clumping together as it
precipitates pulls more strands along with it as it rises into the
alcohol.

You can use a wooden stick or a straw to collect the DNA. If you
want to save your DNA, you can transfer it to a small container filled
with alcohol.

You Have Just Completed DNA (with some RNA) Extraction!

Some other possibilities for investigation

Now that you've successfully extracted DNA from one source, you're ready to experiment further.
Try these ideas or some of your own:
• Experiment with other DNA sources. Which source gives you the most DNA? How can you
compare them?
• Experiment with different soaps and detergents. Do powdered soaps work as well as liquid
detergents? How about shampoo or body scrub?
• Experiment with leaving out or changing steps. We've told you that you need each step, but is
this true? Find out for yourself. Try leaving out a step or changing how much of each ingredient
you use.
• Do only living organisms contain DNA? Try extracting DNA from things that you think might not
have DNA.

And now some questions (and answers!)

1. What is biotechnology ?
Ans. Biotechnology deals with techniques of using live organisms or enzymes from organisms
to produce products and processes useful to humans.

2. What is recombinant DNA ?

Ans. Recombinant DNA is the DNA formed by combining DNAs from two different sources/
organisms.

3. What is genetic engineering ?

Ans. If refers to the techniques which alter the genetic material (DNA or RNA) of host organisms
and thus change the phenotype of the host organisms. (For instance bananas have been
genetically modified in order to create vaccines against cholera and hepatitis!)

4. What are Genetically Modified Organisms (GMOs) ?

Ans. These are the organisms whose genes have been altered by human intervention.

5. Meat tenderizer probably contains the enzyme cellulose. Why is this important in the
extraction of DNA from plant cells ?
Ans. The enzyme cellulase is used to digest the cellulose cell wall present in plant cells.

6. 'What is the role of detergent in isolation of DNA?

Ans. Detergent dissolves the membranes that enclose the DNA within the cell.

7. Why is salt added to see DNA?

Ans. Salt water allows the DNA to precipitate, when alcohol is added to the solution.
(Precipitate happens when something turns into a solid form, which can be seen or collected.)

8. What else does meat tenderizer do in the process of isolation of DNA?

Ans. Meat tenderizer contains enzymes which dissolve the proteins which surround the DNA.

9. After adding chilled ethanol, why should the mixture be allowed to stand for few
minutes?
Ans. It allows more DNA to precipitate in the alcohol layer.

Q.10. This investigation works best in chilled conditions. Why is that?

Ans. Chilling protects the DNA from cellular enzymes and also increases the yield of DNA.

Q.11. Where do you find DNA?

Ans. Nucleus of a cell. (And maybe in the mitochondria and chloroplasts.)

Q.12. What is the full name of DNA?

Ans. Deoxyribose nucleic acid

Q.13. Who first modelled the structure of DNA?

Ans. Nobel prize winners, James Watson and Francis Crick, in 1953, first modelled DNA with
the help of X-ray crystallography images provided by Rosalind Franklin (who didn’t receive the
Nobel Prize!).

Q.14. Name the chemicals ‘building blocks’ which combine to make a DNA molecule
Ans. Deoxyribose sugar, phosphate and nitrogenous bases.

Q.15. How is DNA different from RNA?

Ans. The differences between DNA and RNA are as follows:

DNA
Found in chromosomes of nucleus of
eukaryotic cells and free in the
cytoplasm of prokaryotic cells.
Double-helix structure.
Pentose sugar is deoxyribose.
Contains the base thymine
RNA
Mostly found in cytoplasm.
Single-stranded helix.
Pentose sugar is ribose.
Contains the base uracil

Q.16. What are the building blocks of nucleic acid?

Ans. The nucleotides are the building blocks of nucleic acids. Each nucleotide consists of one
nitrogen containing base attached to a pentose sugar (five carbon) which is in turn attached to a
phosphate group.