INTERNATIONAL MEDICAL SCHOOL

BACHELOR’S IN MEDICAL SCIENCE

SUBJECT/CODE: GENETIC DISORDER/QDS20101P

NAME: ILHAASHINI KRISHNAN

Cells are basic unit of like which make up living organism.Deoxyribonucleic acid(DNA) is a

molecule which control activity of the cells.It contains genetic material,instructions that direct

the activities of the cell.When DNA in nucleus ,it is tightly coiled around a set of proteins called

histones.DNA extraction is the process of isolating DNA from the cells of an organism isolated

from a sample.The detergent solution which can break up the fats and proteins.During DNA

extraction,the detergent breakdown the phospholipid membrane surrounding the cell and

nucleus.The salt enables the DNA strands to aggregate. Salt shields the negative phosphate

ends of DNA that allows the ends to come closer so that DNA could precipitate out of the

alcohol solution.The DNA precipitates out of the solution when the alcohol is added.The

alcohol separates the DNA from the other cell components.DNA is not soluble in alcohol.But

its slightly soluble in cold alcohol. Extraction of DNA from an onion is the simplest and easiest

way to isolate DNA.The onion is used because it has low starch content which allows DNA to

be seen clearly.DNA extraction involves 3 steps.First is the lysis of cell membranes with the

cell wall in some organisms and the membrane of the nucleus to release the

DNA.Secondly,Dissociation from nucleoproteins and denaturation of protein.Then,separation

of DNA from other soluble cellular components. The membranes such as the nuclear

membrane, cell membrane, and cell walls must be broken down in order to bring out the DNA

from an onion cell.

Methods:
1. The onion was chopped
2. A solution consisting of one tablespoon (10 ml) of detergent and 1/4
teaspoon (1.5 g) of table salt was prepared. Distilled water to make a final volume of 100
ml added. The salt was dissolved by stirring slowly to avoid foaming.
3. The chopped onion was added to the solution from step 2.
4. The mixture was heated in a hot water bath at 55-60 C for 10-15 minutes.
5. The mixture was cooled for 5 minutes.
6. The mixture was filtered into the clean test tube.
7. Cold alcohol was added by tilting the test tube at and 45-degree angle and the
alcohol was poured very slowly down the side of the tube.
8. The test tube was placed in its rack and not moved for at least 15 minutes. The
DNA precipitated out immediately between two layers of liquid.
9. After 15 minutes, the DNA was floating on the test tube. A wooden stick was used
to spool it like cotton candy.
Result
Draw the clump made of string-like strands

DNA extracted

The white
ring(cloudy)formation
between the two
layers

Discussion

At first,we chopped the onions into small pieces.This chopping is to break the plant’s cell wall

and cytoplasm to leak out.Then we grind the onion using pestel and mortar.This is type of

physical breakdown of the tough cell walls.The we prepared salt and detergent solution.When

we added salt and detergent mixture to the smashed onions, the detergent helped to

breakdown the phospholipid bilayers of the cell membrane and nuclear membrane and

separate the DNA into solution. Whereas,salt helped create an environment where the

different DNA strands could gather and clump,,making easier to see them.Then we put the

mixed solution into the hot water bath.This hot environment helps to increase the rate of

chemical reactions.Then when we cool the solution for 5 minutes,its decreases the rate of the

chemical reactions.After we add the cold alcohol to the filtered onion liquid,the alcohol will

precipitated the DNA out of the liquid while the rest of the liquid remained in solution.Then we

observed the white DNA strands in the alcohol layer between the two layers.If single DNA we
cannot see in our naked eye.But since the DNA strands are clumped together,therefore we

can see clearly.Then we need to use toothpick to take the DNA like cotton candy.

At the end of our experiment,we successfully observe the white ring(cloudy) formed in the

alcohol layer between the two layers.But we are not able to take out the DNA from the solution

using toothpick.This is due to the stick is too short to be inserted into the test tube.

Otherwise,the volume of the solution is not enough.Therefore the formation of DNA will be

more.There are some precautions need to pay attention in order to get accurate result.Factors

may affect the formation of DNA.We cannot put the solution more than 15 minutes because it

may break down the DNA.We need to stir carefully to avoid bubbles and may shear the fragile

DNA strands.When we add the alcohol the position of the test tube should be in 45 degree

angle in the wall of the test tube.The size of the onions not smashed enough to extract the

DNA.The temperature of water and alcohol may also affect the result.

Exercise
1. Explain the specific purpose of detergent, salt, and alcohol.
In a cell, lysis occurs when membranes are broken apart. Cells have an outer membrane

called the cell membrane. They also have an inner membrane that surrounds the DNA. This

inner membrane is called the nuclear membrane. Cell membranes are made up of two layers

of lipids (fat molecules) and proteins. Chemical lysis is one way to break apart the cell

membrane and nuclear membrane. In chemical lysis, we add a detergent to the cell. The

detergent separates the lipid molecules, causing the membrane to break down. They bind to

grease (lipids), which makes it easier to wash the grease particles off with water.

After the cell and nuclear membranes are broken down, the lipid molecules must be removed.

We add salt solution. This causes the detergent and other cellular debris, such as proteins, to

precipitate. To precipitate means to form a solid that separates from the solution. The DNA

remains dissolved in the liquid solution. It can be removed from the cellular debris by

centrifugation. In centrifugation, the liquid solution spins at high speed so that the precipitate
collects as a pellet at the bottom of a tube. The DNA, which is still dissolved in the liquid, can

be moved to a new sample tube. Alternatively, the precipitate can be filtered out of the solution.

That way, the liquid containing the DNA is left behind.DNA in the nucleus is wrapped around

proteins called histones. This helps organize the DNA into chromosomes. To remove the

histone proteins, a protease can be added. A protease is an enzyme that breaks down

proteins.

The DNA now needs to be removed from the liquid solution. DNA is soluble in water. That

means it can dissolve in water. However, it is not soluble when alcohol and salt are present.

Lab technicians can add ethanol or isopropyl alcohol (rubbing alcohol) so that the DNA clumps

and form a visible white precipitate. It’s important to use cold alcohol because it allows a larger

amount of DNA to be extracted. If the alcohol is too warm, it may cause the DNA to denature,

or break down.

Conclusion
In conclusion, by extracting DNA from onion cells and aggregating them in salty water, adding

soap to catalyze the reaction then adding alcohol to separate the DNA, the extraction of DNA

from onion cells has been successful. However, the environment in which the experiment was

conducted was not perfect. Therefore, some errors might have occurred, such as the size of

the onion pieces used was not identical; the temperature of the hot water and the alcohol might

not have been similar, they also, might not have been at the optimal temperature for the

reactions to occur; finally, the stirring speed might not have been optimal. In order to reduce

the number of errors in the lab, the temperature and amounts of water and alcohol should be

measured, and finally, the stirring method should be more accurate.

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