SAINT MARY’S UNIVERSITY

BAYOMBONG, NUEVA VIZCAYA 3700

SCHOOL OF HEALTH AND NATURAL SCIENCES
MEDICAL LABORATORY SCIENCE DEPARTMENT

Name: AFAN, CYTHEREA MAE A. Year and Section: BSMLS 2B-2

Code: 5298 Instructor: MR. JUSTINE VICENTE

ACTIVITY 11
Deoxyribonucleic Acid (DNA) Isolation

ABSTRACT
Since different plant species are biochemically diverse, a single deoxyribose nucleic acid (DNA) isolation
protocol may not be appropriate. DNA isolation procedures have been modified and standardized on a regular basis. The
majority of plant DNA isolation procedures now in use are modified variants of the CTAB
(hexadecyltrimethylammonium bromide) extraction process. Depending on the plant species and plant component
employed, modifications are frequently made to the concentration of chemicals used during the extraction method.
Understanding the role of each chemical used during the DNA extraction technique (such as CTAB, NaCl, PVP, ethanol,
and isopropanol) will help create or adjust procedures for greater precision. The chemicals used in the CTAB method of
DNA extraction have been reviewed, as well as their possible functions on the highly evolved yet complex to students and
researchers genome.

INTRODUCTION

Nucleic acids are biopolymers composed primarily of nucleosides. Two types of nucleic acids are known to occur in
nature—the deoxyribonucleic acid (DNA) and the ribonucleic acid (RNA). Both functions specifically in the storage and
expression of the genetic code. Various protocols for DNA preparation from various sources of tissues have been published
over the last few decades. The extraction of nucleic acids, particularly the DNA, consists of three steps:
• Disruption of the cell membrane, including the cell wall for some organisms;
• Dissociation from and denaturation of proteins; and
• Separation of DNA molecule from other cellular components.
Plant DNA isolation differs from the most modern and generic techniques used in animal tissue isolation due to the
cellular structure of plant material vis-à-vis that of animal tissues. Plants contain cell walls composed of cellulosic materials
and some other complex polysaccharides, and the degree to which they must be separated from the nucleic acid material
(particularly RNA) depends on the intended use of the nucleic acids. It is noteworthy to mention that primarily due to their
large genome sizes, plants can yield large quantities of nucleic acids. Plants also contain two other genomes: mitochondrial
and plastid. In both terrestrial plants and macroalgae, the plastid DNA genome is a double-stranded DNA circle, which
contains the genes for plastid rRNA, tRNA, and some other proteins. Plant genomes can range from 120-217 kilobases.

OBJECTIVES

• Extract DNA from plant sources.

• Characterize the isolated DNA.

MATERIALS AND EQUIPMENT

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 SAINT MARY’S UNIVERSITY
BAYOMBONG, NUEVA VIZCAYA 3700
SCHOOL OF HEALTH AND NATURAL SCIENCES
MEDICAL LABORATORY SCIENCE DEPARTMENT

• 2 pieces onion (1/2-piece banana may be utilized in the absence of onion)

• Water
• NaCl (Salt)
• Pure dishwashing liquid
• Iced-cold 70% alcohol (transfer 70% alcohol to a clean container and put on a fridge or bowl with ice)
• Transparent sample container
• Cutting board
• Strainer
• Knife
• Teaspoon
• Stove
• Blender or mixer

PROCEDURES

1. Peel the onion and chop into small pieces.

2. Put 30 mL of water in a clean transparent container.
3. Add one teaspoon of salt to the 30 mL water. Stir.
4. Add two teaspoons of dishwashing liquid to the solution. Stir well. Avoid bubble formation.
5. Put the chopped onions in the blender or bowl if mixer will be utilized.
6. Add the prepared solution to the blender or to the bowl.
7. Blend/ mix them for about one to two minutes to get a smooth mixture.
8. Pour the mixture from the blender into a transparent container.
9. Heat the mixture in a very low fire for about 1-2 minutes. Stirring it gently from time to time.
10. Filter the mixture. Gather the filtrate on a clean transparent container.
11. Add about 15-20 mL iced-cold 70% alcohol to the filtrate gently.
12. Observe the DNA strands.

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 SAINT MARY’S UNIVERSITY
BAYOMBONG, NUEVA VIZCAYA 3700
SCHOOL OF HEALTH AND NATURAL SCIENCES
MEDICAL LABORATORY SCIENCE DEPARTMENT

RESULT and DISCUSSION

Add two teaspoons of dishwashing liquid to

the solution. Stir well. Avoid bubble
formation.

Heat the mixture in a very low fire for about

1-2 minutes. Stirring it gently from time to
time

| BIOCHEMISTRY LABORATORY
 SAINT MARY’S UNIVERSITY
BAYOMBONG, NUEVA VIZCAYA 3700
SCHOOL OF HEALTH AND NATURAL SCIENCES
MEDICAL LABORATORY SCIENCE DEPARTMENT

I add about 15-20 mL iced-cold 70% alcohol

to the filtrate gently.

There is formation of cloudy like structure after

putting the iced cold 70% alcohol

DISCUSSION: The onion has a low starch content which can be used to see the DNA clearly upon the experiment. The
salt acts as a barrier for DNA's negative phosphate ends, allowing them to move closer together and precipitate out of a cold
alcohol solution. The cold alcohol helps the DNA precipitate (solidify and appear) more quickly. The detergent and heat
break down the membrane phospholipids and proteins, releasing the cell's DNA.
QUESTIONS
1. What are genes, histone and nucleosomes?
The simplest physical and functional unit of heredity is the gene. DNA is the material that makes up genes. Some
genes serve as instructions for the production of proteins. Many genes, however, do not code for proteins. Genes in
human range in size from a few hundred to over 2 million DNA bases.
A nucleosome is a DNA strand that is wrapped around a protein core. The nucleosome is the most basic component
of chromatin. Each nucleosome is made up of a histone octamer, which is made up of a little less than two rounds
of DNA wrapped around a collection of eight proteins called histones.
2. What are the bases found in a DNA? In an RNA?
- There are four nucleotides, or bases, in DNA: adenine (A), cytosine (C), guanine (G), and thymine (T). These
bases form specific pairs (A with T, and G with C). While RNA there are four nitrogenous bases found in RNA:
adenine, guanine, cytosine, or uracil. Adenine and guanine are known as purine bases while cytosine and uracil
are known as pyrimidine bases
3. What type of intermolecular forces stabilizes the secondary structure of DNA?
The double helical helix of DNA is formed by hydrogen bonding between base pairs. In contrast to covalent and
ionic connections, hydrogen bonds do not trade or share electrons. Hydrogen bonds develop quickly over short
distances and are simple to make and break.
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BAYOMBONG, NUEVA VIZCAYA 3700
SCHOOL OF HEALTH AND NATURAL SCIENCES
MEDICAL LABORATORY SCIENCE DEPARTMENT

4. What is the structural difference between ribose and deoxyribose?

Deoxyribose is the pentose sugar in DNA, and ribose is the sugar in RNA. The presence of the hydroxyl group on
the 2' carbon of ribose and its lack on the 2' carbon of deoxyribose distinguishes the sugars.

CONCLUSION: I successfully extracted DNA from the onion. And see onions DNA without a microscope. In isolating I
struggled I tried it twice to achieve the isolation objectives. This DNA extraction laboratory is an activity to facilitate
learning about cells and the structures inside of them. All living things are made of cells. Inside cells are smaller structures
called organelles that work to perform different functions, or jobs, within the cell.

Sources:
https://medlineplus.gov/genetics/understanding/basics/gene/#:~:text=A%20gene%20is%20the%20basic,more%20than%2
02%20million%20bases.
https://www.news-medical.net/life-sciences/Interactions-That-Hold-DNA-Together.aspx
https://bio.libretexts.org/Bookshelves/Microbiology/Book%3A_Microbiology_(Kaiser)/Unit_7%3A_Microbial_Genetics
_and_Microbial_Metabolism/19%3A_Review_of_Molecular_Genetics/19.6%3A_Ribonucleic_Acid_(RNA)#:~:text=The
re%20are%20four%20nitrogenous%20bases,3).
https://bio.libretexts.org/Courses/University_of_California_Davis/BIS_2A%3A_Introductory_Biology_(Easlon)/Readings
/04.4%3A_Nucleic_Acids#:~:text=The%20pentose%20sugar%20in%20DNA,2'%20carbon%20of%20the%20deoxyribos
e.
https://www.nature.com/scitable/definition/nucleosome-nucleosomes-30/
https://www.youtube.com/watch?v=priDTavoEK4

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