DNA Extraction

Objective:
Perform DNA extraction on selected samples using simple chemicals.
An initial step to do genetic recombination is of course to extract DNA. To isolate nucleic acids from
cells, the cells are opened by enzymatically destroying the macromolecules. It can be done using
detergent solution combined with buffering compounds. In the succeeding exercise/lab activity, you are
expected to extract DNA from the selected organism.
Materials:

• ripe banana • pipet

• 1 toothpick
• 1 resealable zipper freezer bag
• metric measuring spoons
• 100 mL beaker
• 1 g table salt
• 100 mL distilled water
• liquid detergent
• ice bucket with ice • water bath
• 1 sealed test tube containing ice-cold alcohol
Procedures:

1. Cut the peeled banana into small pieces and place in the storage bag. Label the bag and make
sure that it is sealed.
2. Mash the banana by pressing the bag with your hands.
3. Add 100 mL water to the mashed banana and continue mashing until it Is completely liquified.
4. To the solution, add 1 g of salt and 10 mL of liquid detergent. Mix slowly to avoid foaming.
5. When the mixture is thoroughly mixed and liquified, place the bag in a 60 oC water bath for 10
minutes.
6. Wash and dry the beaker then place filter paper about ¼ into it. Fold the edges of the filter down
around the outside lip of the beaker so that it will stay in place when the beaker is set down.
7. After the hot bath is completed, carefully place the bag into the ice bath for five minutes.
8. After the ice bath, slowly pour banana solution on the filter paper and allow it to filter into the
beaker until you have approximately 2 cm of filtrate. This may take several minutes.
9. Dispense 2 mL solution into the test tubes containing the alcohol. Do not shake.
10. Let the tubes sit for two to three minutes without disturbing them. Observe and record what
you see.
11. Gently lower a toothpick into the layer of DNA and slowly move the toothpick in small circles to
spool the DNA. Observe and record your observations of the spooled DNA.
 DNA Extraction

Name: Tristan Manalo, Kier Vincent Salvador, Ken Villamin Date Performed: 04/30/24
Section: STEM ENG 2A Date Submitted: 05/02/24
Group No.: Group 10

Hypothesis:

When extracting DNA from a ripe banana using the provided materials, we notice that the
combination of liquid detergent, table salt, and distilled water will effectively break down the cell
membranes and nuclear envelope, releasing the DNA into the solution. The cold temperature
provided by the ice bucket and ice-cold alcohol will aid in the precipitation of DNA, promoting its
separation from other cellular debris. Additionally, the mechanical disruption caused by the
toothpick will assist in breaking apart the banana cells, facilitating the release of DNA. We
hypothesize that this method will result in a visible, albeit small, quantity of banana DNA extract,
which can be observed in the test tube after centrifugation. The humanized result could be: 'We
expect that by combining the detergent, salt, and distilled water, along with the cold temperatures
and mechanical disruption, we'll coax the banana cells to give up their DNA, which should form a
cloudy layer in the test tube after spinning it down. This might not be a lot of DNA, but it'll be
enough to see and study under the microscope.

Result and Interpretations:

After conducting the DNA extraction experiment using the materials listed, a visible white
stringy substance was observed at the interface between the ice-cold alcohol and the banana
mixture in the test tube. Also, the DNA extraction process successfully yielded visible strands of DNA
from the ripe banana. The banana's cellular structure was disrupted by mashing it in the resealable
zipper freezer bag with distilled water. This allowed the DNA to be released into the solution. The
addition of ice-cold alcohol served to precipitate the DNA out of the solution, forming the visible
strands.

Figure 1:

Short Description of Figure 1:

In this part of the experiment, we already mix all of the needed materials, the small
particles you can see is the DNA that extracted in the ripe banana with the help of, liquid
detergent, ice, denatured alcohol.
 Conclusion:
Using just a ripe banana, some household items, and basic laboratory equipment, we
successfully extracted DNA. By mashing the banana in a resealable bag with distilled water and a
bit of salt, we released the DNA from the cells. Adding detergent helped break down the cell
membranes further. Then, by carefully layering ice-cold alcohol on top of the mixture, we were
able to precipitate the DNA out of solution, forming visible strands.

Guide Questions:
1. Explain the purpose of the following steps:

a. mashing the banana

We are essentially cracking open those cell walls by shattering the banana. It is
comparable to using a hammer to open a treasure vault and extract priceless DNA. The
DNA can more easily be extracted later on because it can mix with the other chemicals in
our mixture after the cells are broken apart.

b. adding the liquid detergent

Adding the liquid detergent helps us break down the barriers that are keeping the
DNA locked up inside the cells, making it easier for us to extract it and learn more about
the genetic secrets hidden within the banana.

c. adding the water

By adding water, we're essentially creating the perfect environment for our DNA to
be released from the cells and start its journey towards becoming visible strands in our
experiment. It's like giving our DNA treasure a comfortable ride out of the banana and
into our mixture, where we can start to uncover its secrets.

d. adding the salt

Adding salt to the mixture, it's like sprinkling breadcrumbs along our path. The salt
helps to create the right conditions for the DNA to come out of hiding. It does this by
neutralizing the charges on the molecules in the mixture, making it easier for the DNA to
stick together and form visible strands.

2. When washing dishes, what works better, warm or cold water? Which do you think will help
the detergent break open the cell, warm or cold temperature?

Well for my experience, I used warm water first to soaked those harden smudged of the
dish that where in the plate. After that I used the normal water that every people use
which is the tap water that coming out in our faucets.

3. What is the purpose of putting you solution in test tubes containing alcohol?

The purpose of it is to protect that solution, because it might get contaminated if it’s not
inside the test tube.

4. Use observation terms to describe what you see in the test tube and describe what the
spooled DNA looks like.

After putting all of the needed materials, we check the final solution by using a flashlight
under the test tube, we see a group of tiny particles and it look like a pack of cells.

5. After DNA is extracted, what experimental procedure will follow? Make a schematic diagram
of procedures for biotechnology.
 DNA Isolation from Your Cheek Cells

Introduction
Body cells contain a nucleus that carries the genetic material in the form of DNA. DNA can be
collected from sloughed-off skin cells, and bodily secretions such as saliva (which contain cheek cells),
blood stains, semen, etc. In a crime laboratory, DNA extracted from these sources can be analyzed and
used to trace a specific individual. This is possible because each individual has a unique sequence of
DNA bases. This is unique sequence of DNA found in particular individual is called the DNA fingerprint.
This is comparable to the thumb print of an individual.
Objective: Isolate DNA strands from human cheek cells.

Materials:

• Salt solution (dissolve 8 grams of salt in 92 mL of distilled water)

• Liquid detergent solution (mix 25 mL liquid detergent to 75 mL distilled water)
• Plastic drinking cup
• Test tube with stopper
• Test tube rack
• Distilled water
• Ethyl alcohol (95% denatured)
• 3 droppers
• Glass stirring rod
Procedure

1. Pour 10 mL distilled water in a clean plastic drinking cup.

2. Before performing this step, to not brush your teeth or gargle with water so as not to interfere
with the procedure. Put the 10 mL of water into your mouth and swirl the water around at least
one minute. A longer period and more violent swirling will cause more cells to sloughed off. Spit
the water back into the plastic cup.
3. In a test tube, add 1 mL (approximately 20 drops) of salt solution.
4. In the same test tube (in step 3), ass around 5mL of the spitted water.
5. Add 1mL (20 drops) of liquid detergent solution. Detergents will break down the lipids and
proteins in the cell membrane, thereby disrupting the cells and releasing the DNA into the salt
solution.
Note: The action of the detergent can be enhanced by placing the test tube in water at 55 OC.
This will also denature the enzymes that might damage DNA.
6. Cover the test tube with a stopper and mix the contents of the tube gently by inverting the test
tube five times. Do not shake the test tube.
7. While holding the test tube in a slight angle (around 45 degrees), slowly add 5 mL of ethyl alcohol
down the side of the test tube so that it forms a layer over the cell lysate mixture (mixture of
disrupted cheek cells)
Note: Ice cold ethyl alcohol might be more effective to be precipitate DNA
8. Gently hold the text tube upright and observe what happens at the interface between the ethyl
alcohol and the cell lysate mixture. Observe the clouds of white strands at the interface. Those
are the DNA strands. DNA is insoluble in ethyl alcohol so it precipitates where the two liquids
meet. Soap bubbles will get trapped in the DNA strands.
9. Collect the DNA strand by placing a clean glass stirring rod in the test tube and turning it in one
direction (spooling). The DNA strands will wind around the glass stirring rod.
10. Carefully lift the DNA with the rod and transfer it into a new test tube containing 1 mL (20 drops)
of ethyl alcohol. Observe the DNA strands floating DNA strands floating in the ethyl alcohol.
 DNA Isolation from Your Cheek Cells

Name: Tristan jan Manalo, Kier Salvador, Ken Villamin Date Performed: 04/30/24
Section: STEM ENG 2A Date Submitted: 05/02/24
Group No.: Group 10

Observations

1. Describe the strands of DNA you have isolated.

Well in our DNA experiment, we see three particles inside of our solution in the test tube

2. Compare the DNA strands you isolated with those of your classmates. Describe the
possible factors that influenced the difference in the observed DNA strands.

In our observation we saw some of our classmates only have one particle of DNA
in their extraction, so it means that it may have a big factor when it comes to the
result of the DNA

3. Draw the setup done during the collection of the DNA strand with the glass stirring rod.

Conclusion

In essence, we're creating the ideal conditions for our DNA to escape the cells and begin
Its journey towards becoming visible strands in our experiment by adding water. It's similar to
providing a smooth transition for our genetic treasure from the banana into our mixture, where
we can begin to unlock its mysteries. This technique gave us a concrete link to our genetic
composition by demonstrating the ease of use and accessibility of isolating DNA from cheek cells.
It emphasized how crucial fundamental laboratory methods are for extracting and seeing DNA,
like cell lysis and precipitation.

Post-Laboratory Questions
1. With the action of detergents learned from this experiment, describe the potential harm to
aquatic organisms of the practice of disposing large amount of detergent into bodies of
water.

It's like a sudden invasion of chaos for the aquatic life that inhabits this lake, such as fish,
frogs, and insects. Surfactants are strong compounds included in detergents that are
intended to dissolve grease and oils. They stretch out and mix with the water as opposed to
simply disappearing as they hit the water.

2. Give a possible scenario on how DNA isolation and analysis can be used.

They therefore choose to examine your genetic composition in greater detail using DNA
isolation and analysis. They utilize methods similar to the ones we used in our experiment to
separate the DNA from your cells from a tiny quantity of your blood or saliva.

Once they obtain your DNA, they can do a number of analyses with it. They may amplify
particular genes they believe are related to your condition using PCR (polymerase chain
reaction) (Gunasegar & Neela, 2021). They might also read the complete genetic code from
your DNA sample by using methods similar to DNA sequencing.
Reference:
https://www.sciencedirect.com/science/article/abs/pii/S0732889321000626