BIOLOGY INVESTIGATORY PROJECT

Year
2025-26

ALTHAMASH CHOUDHARY
ROLL NO :-
SCHOOL:- SNBP INTERNATIONAL CHIKHALI
TOPIC:- EXTRACTION OF DNA FROM
TWO PLANT SOURCES
CERTIFICATE OF AUTHENTICITY

This is to certify that, A l t h a m a s h c h o u d h a r y , of grade XII, S N B P has successfully

completed the research project on the topic ‘‘DNA isolation’’ under the guidance of Ms.
Geeta (Biology Teacher).

This project is absolutely genuine and does not indulge in plagiarism of any kind.

The references taken in making this project have been declared at the end of this Report.

Signature of the Signature of the

Candidate Teacher In-Charge

Signature of the Signature of the

Principal External Examiner
 INDEX

Certificate

Acknowledgement

Introduction

Objective

Procedure

Results

Conclusion

Precautions

Biblography
 ACKNOWLEDGEMENT

I would like to express my sincere gratitude to my biology

teacher, Mrs. Geeta, for her constant guidance, support, and
encouragement throughout this project on “DNA
Extraction.” Her valuable advice and motivation helped me
understand the concepts clearly and complete the practical
successfully. I am also thankful to my school, SNBP School,
for providing the necessary facilities and resources to carry
out this experiment. Lastly, I extend my appreciation to my
classmates and family for their cooperation and moral
support during the completion of this project.
 DNA Isolation from Onion.

I. Introduction:

Scientists are involved with research in finding

out as much as possible about the DNA in
plants and animals. Although DNA was
discovered in the 1950’s, there still remains a
lot to be known about it, especially how it
regulates the workings of the body and how it
is used to determine the corporeal traits that we
all have.

We should always remember that DNA is just a

chemical named deoxyribonucleic acid.
Because it is a chemical, we can do reactions
with it just like we can work with any other
chemical. In this lab, we will use the chemical
properties of DNA to extract it from the cells of
onions.
Nucleic acid isolation usually consists of
three steps : (1) disruption of cell membranes with the cell wall in some organisms and the
membrane of the subcellular nucleus to release the nucleic acids,(2) dissociation from
nucleoproteins and denaturation of protein (3) separation of DNA from other soluble cellular
components.

The purpose of this laboratory is to give you firsthand experience with DNA by isolating it from plant
tissue. You will start with whole onions and end with a relatively pure preparation of DNA, containing
literally millions of genes. Once isolated, the DNA can be stored in alcohol or dried out. It will actually
be possible for you to hold in your hands the key to an organism's development and structure.

II. Objectives

1. To become familiar with the physical properties of DNA by isolating it from living tissue

2. To Learn The Purpose Of Each step in the isolation procedure as it relates to the physical
and biochemical characteristics of the genetic material
 III. Materials:
• Fresh onions
• Detergent
• Distilled water
• Ethanol (95%)
• Graduated Cylinders (10mL and 100mL)
• Knife
• Beaker
• Glass stirring rod

Solutions:

-Lysing buffer1: 1M TRIS - 10% mL SDS

5M EDTA
5M NaCl
Distilled water

IV. Procedure:

1) Dice an onion with a knife. Half an onion should be plenty for this lab. Use a mortar and
pestle to mash the pieces of onion into a pulpy sludge.
2) Adding distilled water, homogenate in a blender for about 40seconds.
3) Transfer the blended onion into a 250mL, using a funnel.
4) Add 1mL of 10% SDS and stir.
5) Heat the solution.
6) Let the mixture cool for 5 minutes.
7) Transfer to a test tube using a 50mL pipette, and then add ethanol (ice cold) slowly
pipetting down the side of the tube.

1
This buffer solution is used in this lab for several reasons. First of all, the saltiness and acidity (pH) of the solution is
very close to that in living things; as a result, the DNA will like to dissolve into this solution. Secondly, the detergent
is added to help break down cell walls in the onion cells. Cell walls in living things are made of long polar molecules
with a “greasy” end and a charged end. Because detergent is used to break apart greasy particles in your clothes, it
will also work to tear apart the “greasy” molecules in cell walls. It will be important that these cell walls break down
in this lab, because inside the cell is where the DNA is.
 V. Observation:

A clear layer of ethanol should form on top of the

onion filtrate. (DNA is not soluble in ice cold ethanol)
therefore DNA precipitates out of the solution
becoming visible as white strings in the ethanol layer.

Why we need to chill the buffer solution:

As important as DNA molecules are to life, they are still extremely fragile, and break apart easily when
removed from cells. To slow down the rate at which the DNA breaks up, we cool down the buffer solution
to near freezing. Chemical reactions always take place slower in cold solutions than in warm ones, because
there is a lot less energy around to make the reaction take place.

Why we need to mash the onion:

What we want to do by mashing the onion is to either break the cell walls (releasing the DNA into the
juice) or at the very least expose the cell walls so the detergent can break them down.

VI. Conclusion:

DNA molecules are highly complicated and essentials compounds in the organism, and their
method of isolation from the rest of the cell is made of indispensable yet simple stages of
breaking the cell membranes, seperationg the DNA from proteins and RNAs and finally
precipitating the DNA molecules. We can actually see the basic source of life after this
experiment.

VII. Questions:

1) Buffer: a mixture of an acid and its conjugate base, which minimizes change in the
acidity of a solution when an acid or base is added to the solution thus maintaining the pH
almost constant.

Precipitate: a solid separated from a solution in which the solid is not soluble.
 Filter:
A porous material through which a liquid or gas is passed in
order to separate the fluid from suspended particulate matter.

Emulsify: to form emulsions (small globules of one liquid in another

liquid with which the first will not mix)

2) The lysing buffer is used to maintain pH and other roles:

EDTA: destabilizes the cell membrane, prevents precipitation of
DNA, inhibits DNAses and binds divalent metal ions that could form
salt with anionic phosphate group of DNA. NaCl: loosens the cell
wall for increase solubility and stability of DNA, releases the plasmid
DNA and sheared cellular DNA, denatures the DNA of the cell.
SDS: anionic detergent, disrupts ionic interaction between proteins.

3)

Na+

Banana

Purpose: To perform and observe the process of DNA extraction with fruits.
Materials:
 Fruit (Banana)
 Blender
 Strainer
 Measuring Cups & Spoons
 Knife
 Test tubes/glass containers
 Stirring Rod/Wooden Stick
 Liquid Detergent
 Water (Warm)
  Salt
 70% Ethyl/Isopropyl Alcohol (Cold)
 Meat Tenderizer (Alternative: Pineapple juice, Contact lens cleaning solution)

Procedure:
A. Blender Insanity
1. Slice one banana into thin slices and place the slices inside the blender.
2. Add in a pinch of salt into the blender for the saline solution. The salt in the saline
solution neutralizes the negative charges on the DNA and thus enables the DNA
strands to stick together and it becomes less soluble in water. It also helps to remove
proteins that are bound to the DNA strands.
3. Add about 1 cup of warm water (about 60°C) into the blender. As much as possible,
the amount of water should be twice the amount of space taken up by the banana
slices. The warm water is used to inactivate the DNase.
4. Close the lid of the blender tightly and blend the mixture on high for about 10-15
seconds.
B. Soapy Soup
5. Strain the soup with the use of the strainer into a container and ensure that you get
most of the soup and remove the larger particles present.
6. Add 2 tablespoons of liquid detergent into the soup and mix. Soaps/detergents contain
sodium laurel sulfate that removes fats and proteins, thus enable them to cut through
cell membranes and release the DNA because the membranes are made up of fats and
proteins.
7. Allow the soup mixture to sit for 5-10 minutes. This is to make sure that the proteins
and lipids that are in the cell membrane are captured and broken apart. This also
releases the DNA strands from the cell and allows us to see the DNA strands much
more clearly.
C. enzyme power
8. Pour the soup mixture into the test tube/container and fill about 1/3 of the tube/glass
container. If there is still leftover soup mixture, place it in another test tube/ glass
container.
9. Add a pinch of meat tenderizer into the mixture. If there is none available, make use of
pineapple juice or contact lens cleaning solution. This serves as the enzyme to be
added into the mixture which breaks down the proteins protecting the DNA strands.
 D..Alcohol Separation
10. Tilt the test tube and pour cold ethyl/isopropyl alcohol into the mixture. Make sure
that you pour the alcohol down the side of the test tube/container and do it slowly.
Pour alcohol into the tube until the amount of alcohol is about the same as the
amount of soup in the tube and it creates a separate layer on top of the soup mixture.
The cold alcohol is used to precipitate the DNA as the strands are very sensitive to
temperature.
11. Let it sit for about 10-15 minutes. As the soup stays at the bottom layer, the DNA
strands will suspend and move to the alcohol layer. Using the stirring rod/wooden
stick, slowly draw out the DNA strands from the soup and alcohol layer and remove it
by spinning the rod slowly for the DNA to attach and wrap around it. The DNA
extraction process is complete.

Possible Problems/Issues:
1. Possibility of overmixing/blending the soup too much which may end up destroying the
DNA strands and making it difficult for me to see and extract them.
2. Failure to properly strain the mixture and remove unnecessary elements (lipids, protein,
etc.) completely which would make it more difficult to perform the extraction.
3. Failure to extract or the DNA strands from the soup mixture and alcohol properly (unable
to gather and extract much DNA strands from the tube; alcohol used is not cold in
temperature).Negatively charged DNA has a double helical structure maintained by hydrogen
bonds. Sodium cations are essential in preserving this structure because hydrogen bonds are lost
in a medium with low ionic forces.
4. Other vegetables and fruits that can be used for this lab are: Tomatoes, cherries, bananas,
pineapples, apples…
5. Some fruits and vegetables contain polysaccarides which make purification of the DNA
more challenging.
 Conclusion

The DNA extraction experiment provided a clear understanding of

how DNA, the molecule of heredity, can be isolated from living
cells. Using simple materials like detergent, salt, and alcohol, I was
able to separate DNA visibly, proving its presence in all living
organisms. This practical experience deepened my knowledge of
molecular biology and the structure and function of DNA. It also
helped me appreciate how DNA extraction is the first step in many
biological studies, including genetic engineering, forensics, and
research. Overall, the experiment was educational, engaging, and
enhanced my scientific skills.
 Precautions

1. Handle all glassware and laboratory equipment carefully to avoid

breakage or injury.
2. Ensure all apparatus are clean and free from contaminants before
starting the experiment.
3. Use fresh biological material for better DNA yield and clarity.
4. Add chemicals like detergent, salt, and alcohol in correct
amounts as instructed.
5. Avoid shaking the mixture vigorously to prevent breaking the
DNA strands.
6. Use cold alcohol for better precipitation of DNA.
7. Wear gloves and a lab coat to maintain hygiene and safety.
8. Dispose of all chemical waste properly after the experiment.
 Bibliography

· NCERT Biology Textbook for Class XII, National Council of Educational Research and Training
(NCERT).

· Practical Manual of Biology, Class XII, NCERT.

· “Molecular Biology of the Gene” by James D. Watson.

· [Link] – National Center for Biotechnology Information.

· [Link] – Biology Online Resources.

· Guidance and reference from Mrs. Geeta, Biology Teacher, SNBP School.