Extraction of DNA from Cheek Cells

Gene Smith
February 29th 2013
INTRODUCTION
DNA, deoxyribonucleic acid, is the genetic material of every living organism
and is found in the nucleus of eukaryotic cells. DNA is often called the
blueprint for life because it contains the necessary information to carry out
all the living processes of the cell (1). The purpose of this lab was to extract
DNA from human cheek cells. The isolated DNA could be used in mapping or
sequencing, PCR, crime scene investigation or other downstream
applications (2).
MATERIALS AND METHODS
To extract DNA from cheek cells, 3 mL of water was added to a labeled, 15
mL tube. The inner lining of the cheek was then chewed for approximately 30
seconds. The 3 mL of water was used to rinse the mouth for 30 seconds,
after which the water and the cheek cells were expelled back into the 15 mL
tube. Using a plastic transfer pipet, 2 mL of lysis buffer was added to the
tube containing the water with the cheek cells. The tube was then capped
and gently inverted 5 times to lyse the cells.
Five drops of protease and salt solution were added to the sample. The tube
was capped and inverted 5 times. The sample was incubated in a water bath
at 50C for 10 minutes. Following incubation, 10 mL of cold ethanol was
slowly added while holding the tube at a 45 angle. The tube was placed
upright at room temperature for five minutes, after which the sample was
inverted 5 times. The DNA extraction protocol and all materials and reagents
were provided by Bio-Rad (2).
RESULTS

Throughout the experiment, numerous observations were made. The water

containing cheek cells was white or cloudy in appearance. Upon addition of
the lysis buffer and protease, the solution became clear. A white precipitate
was visible following the addition of cold ethanol.
DISCUSSION
Extraction of DNA from cells is a relatively straight-forward and simple
process. It is routinely carried out in laboratories and has numerous
downstream applications, including sequencing and PCR.
Only a few reagents were used to extract DNA from cells: lysis buffer,
protease and ethanol. Lysis buffer broke open the cheek cells to expose DNA
and cellular proteins. This explains why the sample in this experiment
became clear upon addition of the lysis buffer. Protease served to
enzymatically break down the proteins in the sample. Maximum activity of
the enzyme was achieved by incubating the sample at 50C. Finally, DNA
became visible as a white precipitate after adding cold ethanol. Extraction of
DNA was successful following this protocol. The DNA sample can now be
purified for use in future experiments.

REFERENCES
1. Russell, Peter J. 2010, iGenetics: A Molecular Approach, San Francisco:
Benjamin Cummings Publishers, 3rd Ed.
2. www.bio-rad.com