Aim: Isolation of DNA from onion cells using household detergents to obtain a pure DNA

sample.
Requirements: Fresh onion, Blender or mortar and pestle, Household detergent (such as dish
soap), Cold ethanol or isopropanol, Centrifuge, Centrifuge tubes, Pipettes, Microcentrifuge
tubes, Water or TE buffer (for dissolving the DNA).
Principle
The principle behind this experiment is to isolate DNA from onion cells using household
detergents. DNA is the genetic material that contains the instructions for the development and
function of all living organisms, including plants such as onions.
The first step in the protocol is to grind the onion in a blender or mortar and pestle to obtain a
homogenate, which breaks down the cell walls and releases the DNA. The detergent is added
to the homogenate to help break down the cell membranes and release the DNA. The
detergent also helps to denature the proteins that may be associated with the DNA and make
it easier to extract.
Next, cold ethanol or isopropanol is added to the mixture to precipitate the DNA. DNA is
insoluble in alcohol and as a result, it will form a white pellet at the bottom of the centrifuge
tube after spinning it. The supernatant, which contains the other cellular components, is
removed leaving the DNA pellet.
Finally, the DNA pellet is washed with 70% ethanol or isopropanol to remove any remaining
detergent or other impurities. The DNA pellet is then dried and dissolved in a small amount
of water or TE buffer, which is a buffer that helps to maintain the pH of the DNA sample and
prevents it from denaturing.
Overall, the principle of this experiment is to break down the onion cells, release the DNA
and separate it from other cellular components using detergent, ethanol, or isopropanol, and
washing it to obtain a pure DNA sample.
Procedure:
Here is a general protocol for isolating DNA from an onion using household detergents:
1. Prepare homogenizing medium: Prepare cell lysis solution by mixing dish washing
liquid (3 tsp), sodium chloride (1/2 spatula), and RO water (100 mL) in a glass
tumbler. Stir well. (The detergent will help to break down the cell walls and
membranes, releasing the DNA)
2. Homogenate: Peel and grate two medium sized onion (~50.0 g) to prepare a
homogenate. Mix homogenizing medium (Lysis solution) and onion homogenate in a
glass tumbler and stir
3. Heat the preparation: Heat content (containing the homogenate suspended in
homogenizing medium) over the hot plate / water bath at 60-70oC for about 5-10
mins with continuous stirring.
4. Cool the preparation: Quickly cool the preparation by placing it on ice bath. Keep
stirring the solution gently to allow even cooling throughout.
 5. Filter the preparation: Filter the homogenate through strainer.
6. Deproteinize the homogenate: Add 4-6 drops of eye lens cleaning solution to the
filtered homogenate and then keep it for 5 mins. This step is actually DNA
purification which can be avoided*
7. Precipitation: Pour the clear solution or the filtered homogenate on a wide glass dish.
Add 20.0 -30.0 mL of ice-cold chilled spirit/ethanol/isopropanol slowly along the side
of the glass dish (use a straw/dropper). A clear layer of ethanol should form on top of
the onion filtrate in petri dish.
8. Keep the solution undisturbed for 3-5 minutes. Bubbles will form, precipitating the
DNA out of solution which becomes visible as the white strings / thread
9. Spooling: Spool out the stringy DNA onto a spoon by slowly rotating the stem in one
direction. Continue to rotate the stem as you move it in large circles through the dish,
until complete DNA settles on it.
10. Wash the DNA with 70% ethanol or isopropanol to remove any remaining detergent
or other impurities.
11. Air dry the DNA pellet for a few minutes or dry it with filter paper and then dissolve
it in a small amount of water or TE buffer.
Precautions:
1. Sterilization: It is important to sterilize all equipment and materials before conducting
the experiment to prevent contamination of the DNA sample.
2. Homogenization: Care should be taken when grinding the onion to obtain a
homogenate to avoid contamination and to preserve the integrity of the DNA. The
blender or mortar and pestle should be clean and free of any contaminants.
3. Cold ethanol or isopropanol: It's important to use cold ethanol or isopropanol to avoid
the DNA denaturation.
4. Centrifugation: Care should be taken when handling the centrifuge and the centrifuge
tubes to avoid injury and to ensure that the DNA pellet is collected accurately.
5. Storage and handling: The DNA sample should be stored in a suitable buffer or water
and handled carefully to avoid degradation or contamination. The sample should be
stored at appropriate temperature (4°C) and be used as soon as possible.

Note: -It's important to use a blender or mortar and pestle that are clean and free of any
contaminants. -It's also important to use cold ethanol or isopropanol to avoid DNA
denaturation. -This is a basic protocol, and the exact amounts of the reagents, time, and
temperature may vary depending on the type of onion and detergent you are using.
It's worth mentioning that this protocol is a simple method and can be used as a
demonstration of DNA isolation, but due to the impurities that may exist in household
detergent, it's not recommended to use it for research purposes.