Biol 107 – Cell and Molecular Biology

1st Semester A.Y. 2021-2022

Name: Cristhel Joy L. Laganson Section: EOP1

DNA EXTRACTION

The saliva is laden with cells, shed from your inner cheeks and mouth lining, which are full of
Deoxyribonucleic acid (DNA). This is a good source of sample to carry out simple DNA extraction method
which can be performed at home. The process of DNA extraction involves three (3) basic steps: Lysis,
Precipitation and Purification. In Lysis, the cell and the nucleus are broken open to release the DNA inside.
This step uses detergents and enzymes such as Proteinase K to free the DNA and dissolve cellular proteins.
The precipitation step separates DNA from the cellular debris. First, Na+ ions (sodium) neutralize the
negative charges on the DNA molecules, which makes them more stable and less water soluble. Next,
alcohol (such as ethanol or isopropanol) is added and causes the DNA to precipitate out of the aqueous
solution because it is not soluble in alcohol. Finally, purification involves rinsing with alcohol to remove
any remaining unwanted material and cellular debris. At this point the purified DNA is usually re-dissolved
in water for easy handling and storage. In this activity, you will be able to extract your own DNA using the
available resources in your homes. You need to document the whole process through a short video clip
as part of the completion requirements of this activity.

Objectives:

1. To perform extraction of own DNA from saliva;

2. To make use of common household materials for DNA extraction;
3. To describe the principle behind each step of the extraction process.

Materials
• A clean shot glass or any small glass container (not plastic)
• Saliva
• Dishwashing soap (concentrated)
• Table salt
• Pineapple juice (or meat tenderizer, or cleaning solution for contact lenses)
• Chilled alcohol (70% Ethyl or Isopropyl Alcohol)
• Drinking straw or medicine dropper
• Toothpick
Instructions

1. Spit into the shot glass until it’s a one-fourth full of your saliva.
2. Add a few drops of dishwashing soap. This will break open the cells, a process called
“lysing.”
3. Add a small volume of pineapple juice by using a spoon. This will clean up some of the
proteins that have spilled out of your cells alongside your DNA.
4. Add a pinch of salt. This causes the DNA to start clumping together.
5. Swirl the glass to mix the ingredients.
6. Gently fill the rest of the shot glass with high concentration of chilled alcohol, which
should sit in a layer on top of the mixture. To prevent it from mixing too much, you can
add the alcohol gradually with a drinking straw: Put the straw into the alcohol, cap it
with your finger, hold the straw right over the liquid in the shot glass, and release.
Another option is to use a dropper and gradually add it right over the mixture in the
shot glass.
7. Use a toothpick to spool up the cloudy, snot-like material that has formed in the glass.
This is your DNA.

Questions

1. Fill-in the table:

Procedure
1. Addition of dishwashing soap
2. Addition of pineapple juice
3. Addition of salt
Principle/Purpose
The addition of dishwashing soap breaks
down your cheek cell membranes.
Dishwashing soap will also cut through
cell walls because the membranes
surrounding the cells are made up of fats
and proteins. The dish soap pulls apart
the membranes, releasing the DNA. This
causes the DNA to be released into the
salt water.
The addition of pineapple juice helps
untangle and unfold the DNA from the
other parts of the cell.
We can help neutralize the DNA charge
and make the molecule less hydrophilic,
which means it's less soluble in water, by
adding salt. The salt also helps to remove

4. Swirling of the glass
5. Addition of chilled alcohol
proteins that are bound to the DNA and
to keep the proteins dissolved in the
water.
to mix the ingredients
The addition of chilled alcohol helps the
DNA precipitate (solidify and appear)
more quickly.

2. What do you think will happen if salt is not added to the mixture?
If the salt is not added to the mixture, it would not be possible or harder to create an
environment where the different DNA strands could gather and clump, making it harder
for us to see them.
3. Why must alcohol be chilled prior to addition?
It is because chilled alcohol will increase your yield of DNA. The cold alcohol helps the
DNA precipitate (solidify and appear) more quickly. If the alcohol is too warm, it may
cause the DNA to denature [bold], or break down.
4. Why must alcohol be added slowly into the mixture?
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.
5. Why is saliva a better source of DNA for extraction over other body parts like the skin or
scalp?
According to the study of Francini et al., 2017 Saliva when compared to blood collection
has the following advantages: it requires no specialized personnel for collection, allows
for remote collection by the patient, is painless, well accepted by participants, has
decreased risks of disease transmission, does not clot, can be frozen before DNA
extraction and possibly has a longer storage time. Also, it is an excellent source of
testable genetic material and a large amount of DNA can be collected in a relatively
small sample of saliva.

VIDEO LINK:
Google Drive:

https://drive.google.com/file/d/1KssValTijsGNryaL7UDKrkCKswp_0URk/view?usp=sharing

YouTube Link:

https://www.youtube.com/watch?v=yOfxhXXhzoY