EXTRACTION AND ISOLATION OF GENETIC MATERIAL FROM

CHEEK CELLS
JONEL MARK A. CARANDANG, FIEL CRIS A. LANSANG, ELMER P. IBAÑEZ JR., MA.
LOURDES D. HERNANDEZ, KENN S. RUBIS
Abstract:
The present study aimed to determine and observe the appearance of an isolated DNA or
genetic material. It also further aims to identify the significance of DNA extraction to scientists.
Necessary to the experimental study are clear cups, a soap solution with a 25% volume per
volume concentration, stirrer, salt solution with a quarter teaspoon salt in 50mL of water, beaker,
graduated cylinder, cold 70% isopropyl alcohol, water, and protective laboratory clothing.
Initially, the salt and soap solution is separately prepared in beakers. Then, each member was
instructed to gargle the salt solution for a minute and was asked to spit it back to the cup.
Subsequently, 10mL of the dilute soap solution was added to the cheek cells extract in the salt
solution and was then mixed with the use of a stirrer. Next, cold alcohol, 20mL in volume, was
added to the mixture slowly in the side. Finally, the DNA extract was observed and the data
and/or observations were recorded. Hence, it was concluded that the isolation of DNA can be
performed using diluted soap solution, salt, water, and extracted cheek cells. The experimental
study further concludes that the physical appearance of the genetic material can be described a
tiny thread-like structure interconnected to each other
Keywords: DNA, extraction, isolation, genetic material, cheek cells

Introduction development, functioning, growth and

reproduction of all known organisms and
many viruses. DNA and ribonucleic acid
One of the key mystery of the natural (RNA) are nucleic acids; alongside proteins,
world is the mechanism of how organism lipids and complex carbohydrates
thrive and evolve over different arcs of time. (polysaccharides), nucleic acids are one of
In the distant past, mankind has little to none the four major types of macromolecules that
knowledge about the science of the human are essential for all known forms of life.
body. Then came the discovery of
microscope that pave the way to the The two DNA strands are also known
discovery and study of cells and as polynucleotides as they are composed of
microbiology. Later studies determined the simpler monomeric units called nucleotides
factor that enables traits to be passed from (Johnson et. al, 2014) Each nucleotide is
parent and offspring--- the very concept of composed of one of four nitrogen-containing
heredity. One of the greatest discovery of nitrogen bases (cytosine [C], guanine [G],
man is the concept of gene make-up that adenine [A] or thymine [T]), a sugar called
instructs the totality of an organism. The way deoxyribose, and a phosphate group. The
an organism should appear, how his organs nucleotides are joined to one another in a
and organ systems function, and how an chain by covalent bonds between the sugar of
organism passed on these traits to the next one nucleotide and the phosphate of the next,
generation to ensure survival and continuity resulting in an alternating sugar-phosphate
of the race and species in which the organism backbone. The nitrogenous bases of the two
is belonged to. separate polynucleotide strands are bound
together, according to base pairing rules (A
Deoxyribonucleic (DNA) is a with T and C with G), with hydrogen bonds
molecule composed of two chains that coil to make double-stranded DNA. The
around each other to form a double helix complementary nitrogenous bases are
carrying genetic instructions for the divided into two groups, pyrimidines and

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purines. In DNA, the pyrimidines are within individuals over time influence the
thymine and cytosine; the purines are adenine maintenance of health and risk to disease.
and guanine. Human DNA has around 3 Such studies require a detailed understanding
billion bases, and more than 99 percent of of the microbial diversity found at various
those bases are the same in all people, anatomically distinct sites of the human
according to the U.S. National Library of body.
Medicine (NLM).
There are various types of methods to extract
DNA was first observed by a German DNA from organisms. The cultivation-
biochemist named Frederich Miescher in dependent methods commonly used in
clinical and research laboratories have
1869. But for many years, researchers did not
provided a valuable but incomplete picture of
realize the importance of this molecule. It the vast diversity found in the human
was not until 1953 that James Watson, microbiome because many, if not most
Francis Crick, Maurice Wilkins and Rosalind human-associated microorganisms have not
Franklin figured out the structure of DNA — yet been successfully cultured in the
a double helix — which they realized could laboratory (Aas et. al, 2005), (Bik et. al,
carry biological information. 2006), (Pei et. al, 2004), (Zhou et. al, 2004).
These methods are also limited because most
Watson, Crick and Wilkins were do not lend themselves to the analysis of large
awarded the Nobel Prize in Medicine in 1962 numbers of samples because they are labor-
"for their discoveries concerning the intensive and costly.
molecular structure of nucleic acids and its However, the application of cultivation-
significance for information transfer in living independent molecular approaches based on
material." Franklin was not included in the the phylogenetic analysis of the 16S rRNA
award, although her work was integral to the gene sequences provides a way to access the
uncultured majority (Robinson et. al, 2010),
research.
(Ward et. al, 1990), allowing for more
DNA extraction is the very comprehensive comparative studies of
microbial communities associated with the
foundation in which specialists are able to
human body (Eckburg et. al, 2005), (Gao et.
alter or correct sequences. This bear the al, 2007), (Ravel et. al, 2010). Of the several
greatest significance in genetic engineering, rapid and inexpensive DNA isolation
biotechnology, and also in the field of procedures that have been described recently,
medicine and health. DNA extraction is an one of the most popular is that of Saghai-
essential step in all cultivation-independent Maroof et ale (1984), a procedure using
approaches to characterize microbial hexadecyltrimethylammonium bromide
(CTAB) and lyophilized tissue.
diversity, including that associated with the
human body. Various cultivation-independent approaches
to characterizing diversity in microbial
The microorganisms that colonize various communities all require extraction of
anatomical sites of the human body play genomic DNA from the samples of interest.
important roles in human health and disease Previous studies have shown that differences
(Alberts et. al, 2014). For example, bacteria in the structures of bacterial cell walls cause
in the human intestine contribute to digestion bacterial cell lysis to be more or less efficient
of inaccessible compounds (Backhed et. al, (Carrigg et. al, 2007), (Frostegard et. al,
2004) and development of the host immune 1999), (Krsek & Wellington, 1999). This can
system (Cebra JJ, 1999), (Round & distort the apparent composition of microbial
Mazmanian, 2009), while vaginal microbiota communities (Carrigg et. al, 2007), (Morgan
helps prevent urogenital diseases and et. al, 2010), (Salonen et. al, 2010),
maintain health in women (Lai et. al, 2009), (Ariefdjohan et. al, 2010), (Scupham et. al,
(Taha et. al, 1998), (Watts et. al, 2005). 2007), (Inceoglu et. al, 2010) and introduce
In recent years there has been bias in estimates of relative abundances of
microbes in samples (Carrigg et. al, 2007),
increasing interest in knowing more about
(Krsek & Wellington, 1999), (Burgmann et.
how differences between individuals, or
2|Page
al, 2001). However, despite the critical nature ii. To isolate and observe the genetic
of this first step, the selection of a suitable material.
procedure for the extraction of DNA from
human samples has not received enough iii. To explain the principles of DNA
attention (Frostegard et. al, 1999), (Forney et. isolation; and,
al, 2004). Indeed, in many previous
investigations of the human microbiome, the iv. To explain the importance of DNA
genomic DNA extraction methods used were extraction to scientists.
chosen without an obvious rationale, and
used without validation. The present study implies to perform
the procedure to extract DNA from cheek
cells and isolate it. It is fully hoped that the
Multiple criteria, including DNA yield, present experimental study shall provide
DNA shearing, reproducibility, and necessary information on what the DNA
representativeness can be used to evaluate looks like and how each variable led to the
DNA extraction methods. Numerous extraction and isolation of the genetic
investigators have tried to increase the DNA material from cheek cells.
yield through use of physical disruption
methods such as bead beating and sonication
to improve the lysis of bacterial cells. Materials and Method
However, such treatments can shear
genomic DNA into small fragments and this
may lead to the formation of chimeric To perform efficiently, correctly and
products during PCR amplification of gene to arrive with accurate results, the necessary
targets (Liesack et. al, 1991), materials and equipment were gathered,
(Wintzingerode et. al, 1997). In addition, it obtained, and sanitized. Necessary to the
is important to assess the variation between experimental study are clear cups, a soap
analysts and over time. This is especially solution with a 25% volume per volume
important when trying to track differences concentration, stirrer, salt solution with a
across sampling sites, time scales or quarter teaspoon salt in 50mL of water,
treatments, and to compare results obtained beaker, graduated cylinder, cold 70%
by different laboratories. But achieving an isopropyl alcohol, water, and protective
accurate representation of bacterial profiles laboratory clothing. Initially, the salt and
is arguably the most critical criterion soap solution is separately prepared in
(Turnbaugh et. al, 2007), (Peterson et. al, beakers. Then, each member was instructed
2009), because ultimately the objective is to to gargle the salt solution for a minute and
obtain DNA that fairly represents the was asked to spit it back to the cup.
microbial diversity in samples with the least
bias for composition and abundance.
Unfortunately, most studies have evaluated
the efficacy of different DNA extraction
methods using environmental samples
comprised of unknown microbes (Carrigg et.
al, 2007), (Bertrand et. al, 2005), (McOrist
& Jackson, 2002), which make evaluation of
representativeness impossible.
With these considerations in mind,
the present experimental study was aimed to
examine,

i. The DNA from cheek cells and describe its

appearance.

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Figure 1.1 Preparation of the Salt Mixture will not induce a mixing process. It is also
predicted that the DNA would appear to be
very small strand-like structures as described
by books and scientific models.
The said projection was tested by
extracting the genetic material from an
individual’s cheek sell and isolate with the
utilization of salt solution, a diluted soap
solution, and cold isopropyl alcohol.
After pouring the cold alcohol to the
mentioned mixture of gargled salt solution
and diluted soap, tiny thread-like figures
appeared on the upper segment of the
solution and it was determined as the isolated
DNA from the cheek cells.
Figure 1.2 Designated Salt solution for each
member of the experimental study group.

Figure 1.3 Gargling of the salt solution

Subsequently, 10mL of the dilute

soap solution was added to the cheek cells
extract in the salt solution and was then
mixed with the use of a stirrer. Next, cold Figure 2.1 and 2.2 Isolated DNA from cheek
alcohol, 20mL in volume, was added to the cells
mixture slowly in the side. Finally, the DNA
extract was observed and the data and/or The Figure exhibits small traces of thread-
observations were recorded. like figures that is the isolated DNA or
genetic material from the cheek cells.

Results and Discussion:

The initial prediction of the study is
that the genetic material will be exposed in
the segment of the alcohol due to which there
is a difference in the property with water that

4|Page
 why a diluted soap solution is used in the
process of DNA isolation.
The initial role of the ethanol and
monovalent cations is to remove the
solvation shell surrounding the DNA and
permitting the precipitation of the DNA in
pellet form. The ethanol also serves to
promote the aggregation of the DNA. With
respect to the washing steps, typically a 70%
ethanol solution is used. This permits the
solubilisation of the salts whilst minimising
Figure 2.3 All of the isolated DNA from the the solubility of the DNA. The salts are
experimental group. therefore removed due to solubility
differences, especially with the aggregated
Nuffield Foundation (2011), explains
DNA. The final 100% ethanol wash that is
that using ice-cold ethanol and ice-cold water
usually employed serves more to permit the
increases the yield of DNA. Low
easier evaporation of the ethanol from the
temperatures protect the DNA by slowing
DNA pellet in order to prevent any carry
down the activity of enzymes that could
over.
break it apart. A cell’s DNA is usually
protected from such enzymes (DNases) by DNA is less soluble in isopropanol so
the nuclear membrane which is disrupted by it will fall out of solution faster and at a lower
adding detergent. DNases in the cytoplasm concentration, but the downside is that the
would destroy the DNA of viruses entering salt will too. With ethanol, the DNA needs to
the cell. Cold ethanol helps the DNA to be at a higher concentration to flocculate but
precipitate more quickly. Chill the ethanol in the salt tends to stay soluble, even at cold
a screwcap plastic bottle in the prep room temperatures.
freezer. Below 4 °C ethanol is below its
DNA falls out of solution in 35%
flashpoint so this is safe even if your freezer
isopropanol and 0.5M salt. Using ethanol, the
is not spark proof. This further justify the
final concentration needs to be around 75%
usage of cold isopropyl alcohol in the
with 0.5M salt. So for the typical
experimental design.
precipitation protocol, isopropanol is added
The isolation of DNA usually begins from between 0.7-1 volumes of sample and
with lysis, or breakdown, of tissue or cells. ethanol is added at 2-2.5 volumes of sample.
This process is essential for the destruction of
DNA extraction is beneficial to
protein structures and allows for release of
scientist in the field of genetic engineering,
nucleic acids from the nucleus. Lysis is
medicine, and in biotechnology producing
carried out in a salt solution, containing
genetically modified organisms to heighten
detergents to denature proteins or proteases
their efficiency and even cute defects in the
(enzymes digesting proteins), such as
gene sequence of an organism.
Proteinase K, or in some cases both. It results
in the breakdown of cells and dissolving of One of the earliest discovery was of
membranes. the study conducted by Pääbo (1989). In the
study, several chemical and enzymatic
ResearchGate (2017), determined
properties were examined in the DNA
that detergent contains sodium laurel sulfate,
extracted from dry remains of soft tissues that
which cleans dishes by removing fats and
vary in age from 4 to 13,000 years and
proteins. It acts the same way in the DNA
represent four species, including two extinct
extraction protocol, pulling apart the lipids
animals (the marsupial wolf and giant ground
and proteins that make up the membranes
sloth). The DNA obtained was invariably of
surrounding the cell and nucleus. Once these
a low average molecular size and damaged by
membranes are broken apart, the DNA is
oxidative processes, which primarily
released from the cell. This was the reason

5|Page
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Conclusion Cebra JJ (1999) Influences of microbiota on

intestinal immune system development. Am
It was concluded in the present study J Clin Nutr 69: 1046S–1051S.
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Dethlefsen L, McFall-Ngai M, Relman DA
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appearance of the genetic material can be
described a tiny thread-like structure Eckburg PB, Bik EM, Bernstein CN, Purdom
interconnected to each other. E, Dethlefsen L, et al. (2005) Diversity of the
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