CERTIFICATE

Date: __________
This is to certify that this work entitled MUTATION was done under my
guidance and submitted by M.krishnakanth, (Regn. No.

) for All

India Senior Secondary Practical Examination 2017 is the original work of

the candidate.

Signature of the
Candidate

Signature of the
Internal Examiner

Signature of the
Principal

Signature of the
External Examiner

ACKNOWLEDGEMENT
First of all, I thank God, almighty who has blessed me and given
this opportunity to submit my biology project.
Secondly, I would like to express my special thanks of gratitude to
my teacher Mrs. Raja Lakshmi M.SC., B.ED., M.PHIL., as well
as our principal ,who gave me the golden opportunity to do this
wonderful project on the topic Extraction Of Onion DNA which
also helped me in doing a lot of experiment and I came to know
about so many new things I am really thankful to them.
Next, I would also like to thank my parents, sisters, brother and
friends who helped me a lot in finalizing this project within the
limited time frame.
M.krishnakanth , X11 A

CONTENTS

Introduction
Benefits of DNA extraction
Theory
Materials required
Procedure
Observation
Conclusion
Precautions
Inference

INTRODUCTION:
Deoxyribonucleic acid (DNA) is a molecule that encodes
the genetic instructions used in the development and functioning of all known
living organisms and many viruses. Along with RNA and proteins, DNA is one

of the three major macromolecules essential for all known forms of life. Most
DNA molecules are double-stranded helices, consisting of two
long biopolymers of simpler units called nucleotides. DNA is well-suited for
biological information storage, since the DNA backbone is resistant to cleavage
and the double-stranded structure provides the molecule with a built-in
duplicate of the encoded information. Many methods have been developed to
purify DNA from organisms, such as phenol-chloroform extraction, and to
manipulate it in the laboratory, such as restriction digests and the polymerase
chain reaction. Modern biology and biochemistry make intensive use of these
techniques in recombinant DNA technology. Recombinant DNA is a man-made
DNA sequence that has been assembled from other DNA sequences. They can
be transformed into organisms in the form of plasmids or in the appropriate
format, by using a viral vector. The genetically modified organisms produced
can be used to produce products such as recombinant proteins, used in medical
research, or be grown in agriculture.

BENEFITS OF DNA EXTRACTION:

DNA extraction serves many different purposes. Evidence stemming from
DNA tests has convicted or exonerated numerous people in murder and other
criminal cases. It's also the deciding factor in many paternity cases. Tests

have also been able to unlock hereditary traits and ancestral paths. Testing
has helped identify bodies found that can't be verified any other way.

THEORY:
The process of extracting DNA from a cell is the first step for many
laboratory procedures in biotechnology. The scientist must be able to
separate DNA from the unwanted substances of the cell gently enough so
that the DNA is not broken up. An onion is used because it has a low starch
content, which allows the DNA to be seen clearly. The salt shields the
negative phosphate ends of DNA, which allows the ends to come closer so
the DNA can precipitate out of a cold alcohol solution. The detergent causes
the cell membrane to break down by dissolving the lipids and proteins of the
cell and disrupting the bonds that hold the cell membrane together. The
detergent then forms complexes with these lipids and proteins, causing them
to precipitate out of solution.

MATERIALS REQUIRED:

two measuring cups (1000 ml) with ml markings

one measuring cup (250 ml) with ml markings
measuring spoons
sharp knife for cutting onion
large spoon for mixing

food processor or blender

thermometer that will measure 600 C (1400 F), such as a candy
thermometer
strainer or funnel that will fit in a measuring cup
coffee filter or cheese cloth
hot tap water bath (600 C)
ice water bath
distilled water
clear-colored shampoo, such as Suave Daily Clarifying Shampoo
large onion
table salt, either iodized or non-iodized
Pasteur pipettes or medicine droppers
95% ethanol (grain alcohol)

PROCEDURE:
Set up hot water bath at 55-60o C and an ice water bath.
For each onion, make a solution consisting of one tablespoon (10 ml)
of liquid dishwashing detergent or shampoo and one level 1/4
teaspoon (1.5 g) of table salt. Put in a measuring cup of 250 ml beaker.
Add distilled water to make a final volume of 100 ml. dissolve the salt
by stirring slowly to avoid foaming.
Coarsely chop one large onion with a knife and put into a measuring
cup (1000 ml). For best results, do not chop the onion too finely. The
size of the pieces should be like those used in making spaghetti. It is
better to have the pieces too large than too small.
Cover chopped onion with the 100 ml of solution from step 2.
The detergent dissolves the fatty molecules that hold the cell

membranes together, which releases the DNA into the solution. The
detergent, combined with the heat treatment used in step 5, causes
lipids (fatty molecules) and proteins to precipitate out of the solution,
leaving the DNA. The salt enables the DNA strands to come together.
Put the measuring cup in a hot water bath at 55-60 C for 10-12
minutes. During this time, press the chopped onion mixture against the
side of the measuring cup with the back of the spoon. Do not keep the
mixture in the hot water bath for more than 15 minutes because the
DNA will begin to break down.
Cool the mixture in an ice water bath for 5 minutes. During this time,
press the chopped onion mixture against the side of the measuring cup
with the back of the spoon. This step slows the breakdown of DNA.
Filter the mixture through a coffee filter or four layers of cheese cloth
placed in a strainer over a measuring cup. When pouring the mixture
into the strainer, avoid letting foam get into the measuring cup. It can
take more than an hour to recover most of the liquid.
Add cold alcohol to the test tube to create an alcohol layer on top of
about 1 cm. For best results, the alcohol should be as cold as possible.
The alcohol can be added to the solution in at least three ways. (a) Fill
a Pasteur pipette with alcohol, put it to bottom of the test tube, and
release the alcohol. (B) Put about 1 cm of alcohol into the bottom of a
test tube and add the onion solution. (c) Slowly pour the alcohol down
the inside of the test tube with a Pasteur pipette or medicine

dropper. DNA is not soluble in alcohol. When alcohol is added to the

mixture, all the components of the mixture, except for DNA, stay in
solution while the DNA precipitates out into the alcohol layer.

Let the solution sit for 2-3 minutes without disturbing it. It is
important not to shake the test tube. You can watch the white DNA
precipitate out into the alcohol layer. When good results are obtained,
there will be enough DNA to spool on to a glass rod, a Pasteur pipette
that has been heated at the tip to form a hook, or similar device. DNA
has the appearance of white mucus.

OBSERVATIONS:
The DNA will slowly rise from the watery lower layer up into the alcohol
layer above it. The DNA will look stringy and have small bubbles attached to
it. It will be a clear, snotty substance, and may be hard to see. Slowly twist
substance onto a toothpick.

CONCLUSION:
DNA is finally extracted from the onion solution. DNA is an actual thing,
found in our bodies, in very large quantities. In fact, it is found in every living
thing except bacteria. It is DNA that determines who we are, but DNA is made
up of the same basic materials as are found in the rest of bodies. It has the
biochemical property of being polar, which allows us to separate it from a
solution using the electrochemical properties of other things found in our
bodies, such as proteins. DNA is important in several other branches of science,
such as genetic engineering.

PRECAUTIONS:
Caution should be taken when the students are chopping the onion
and using the blender. Also, goggles are recommended when
Chopping the onion.
Gloves should be worn at all times to prevent contamination of the
onion sample with nucleases that naturally occur on the skin. Care
should be taken not to touch the glassware or utensils with bare hands
as contamination can occur here as well.
Onion should not be chopped too small as DNA breaks.
Onion solution should not be warmed more than 15min in 550 C.

REFERENCE:

http://www.biotech.iastate.edu/lab_protocols/DNA_Extraction
_Onion.html
http://dwb4.unl.edu/Chem/CHEM869N/CHEM869NLinks/cpmc
net.columbia.edu/dept/physio/tchrplan/oniondna.html
www.wikepidia.org