Catanduanes State University

COLLEGE OF SCIENCE
Natural Science Department
Virac, Catanduanes

Activity 4
Isolation and Characterization of Nucleic Acids

Objectives
At the end of this chapter, the students should be able to:
1. Isolate DNA from microbial, plant, and animal sources;
2. Determine the purity of isolated DNA; and
3. Analyze how biological traits are passed on to successive generations.

Introduction
Deoxyribonucleic acid, or commonly called DNA, is a stranded molecule that determines
our identities and presents in all living organisms including plants, animals and bacteria. In eukaryotic
organisms, like us and vegetables we eat, DNA strands are stored in a cellular structure called nucleus.
However, unlike other cellular structures, it is not easy to visualize the DNA strands in the nucleus
because they are inside of nucleus and very closely packed together. In order for us to see these DNA
strands, we need to open up the cell and unpack them into long chains. Mechanical force is enough to
break clumps of cells into individual cells, but such force may not be enough to break down some cellular
structure as well as DNA packing materials. Strands of DNA only can be separated when all the
supporting materials are chemically broken down, hence releasing thread-like DNA strands.
DNA is made up of building blocks known as nucleotides. Each nucleotide is composed of three
parts—a phosphate group, deoxyribose sugar, and one of the four nitrogenous bases: adenine, guanine,
cytosine, or thymine. The two strands of DNA are the backbone of the ladder, made of carbohydrate
sugar phosphodiester groups. The sugar backbone acts as a support for the rungs of the ladder. The rungs
are composed of the nitrogenous bases. Scientists use the first letters of these bases—A, G, C, and T—to
designate the order of the bases within the DNA strands. The bases are always arranged in pairs. When A
occurs on one strand, T will occur on the opposite strand. Similarly, G and C are base pairs on opposite
DNA strands. The bases are held together by weak hydrogen bonds.
DNA plays an important role in two processes.
During the process of replication, DNA provides information to
copy itself, so genetic information can be passed on from
one generation of cells to the next. DNA also provides
instructions for making proteins, and these instructions are
vital to the maintenance and function of cells. DNA provides the
information on how to order the amino acids required for
making various proteins.

DNA Isolation from Banana

Materials

o DNA source: banana

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 Catanduanes State University
COLLEGE OF SCIENCE
Natural Science Department
Virac, Catanduanes
o 2 spoons (or 30ml) Liquid detergent
o Pineapple juice, papaya juice, or meat tenderizer
o 70%-95% alcohol
o Graduated cylinder or any measuring devices
o Ziplock bags or ice bags
o 250 ml Beaker/Small transparent plastic cups
o 100 ml Cold water
o ½ tablespoon Table salt
o Extra spoon

Procedure

1. Obtain 1/2 cup of material (Banana), and mash the material inside a plastic bag. Mashing will
provide mechanical force to separate cells into individual cells.
2. Transfer the mashed material into a transparent plastic cup and add 100 mL of cold water, until
the mash has milk shake consistency, and ½ spoon of table salt. Mix the content until all salt
crystals are dissolved.
3. Add 2 spoons of liquid detergent (~30 ml) and swirl to mix. In this step, the liquid detergent will
act as a surfactant that disrupts integrity of the cell membrane, therefore, exposing intracellular
materials.
4. Add a small amount of pineapple juice, papaya juice, or meat tenderizer (1 spoon) to the plastic
cup. And swirl the cup very gently. Vigorous stirring is not desired because it may break DNA
strands into smaller pieces, making it harder to see.
5. Tilt the cup and very carefully and slowly pour alcohol into the cup. Add alcohol until you have
about the same amount as the mixture. Do NOT stir the mixture too much. The reason for adding
alcohol is to separate DNA molecules normally dissolved in water. When salty DNA comes in
contact with alcohol, it becomes undissolved, therefore it precipitates out into alcohol layer.

Observation and Result:

Magnification: _____________________

DNA Isolation from Chicken Liver

Materials

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 Catanduanes State University
COLLEGE OF SCIENCE
Natural Science Department
Virac, Catanduanes
o DNA source: chicken liver
o 2 spoons (or 30ml) Liquid detergent
o Pineapple juice, papaya juice, or meat tenderizer
o 70%-95% alcohol
o Graduated cylinder or any measuring devices
o Ziplock bags or ice bags
o 250 ml Beaker/Small transparent plastic cups
o 100 ml Cold water
o ½ tablespoon Table salt
o Extra spoon
o Blender

Procedure
1. Obtain 1/2 cup of material (Chicken liver), and homogenize the material using blender.
2. Transfer the homogenized material into a transparent plastic cup and add 100 mL of cold water
and ½ spoon of table salt. Mix the content until all salt crystals are dissolved.
3. Add 2 spoons of liquid detergent (~30 ml) and swirl to mix. In this step, the liquid detergent will
act as a surfactant that disrupts integrity of the cell membrane, therefore, exposing intracellular
materials.
4. Add a small amount of pineapple juice, papaya juice, or meat tenderizer (1 spoon) to the plastic
cup. And swirl the cup very gently. Vigorous stirring is not desired because it may break DNA
strands into smaller pieces, making it harder to see.
5. Tilt the cup and very carefully and slowly pour alcohol into the cup. Add alcohol until you have
about the same amount as the mixture. Do NOT stir the mixture too much. The reason for adding
alcohol is to separate DNA molecules normally dissolved in water. When salty DNA comes in
contact with alcohol, it becomes undissolved, therefore it precipitates out into alcohol layer.

Magnification: _____________________

Name of Student : _____________________________ Date Submitted : _____________

Course/Year/Block : _____________________________ Instructor : _____________

Activity 4
Isolation and Characterization of Nucleic Acids

DATA SHEETS

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 Catanduanes State University
COLLEGE OF SCIENCE
Natural Science Department
Virac, Catanduanes

1. Explain why the following materials are used in the isolation of DNA:

a. Banana
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________

b. Chicken Liver
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________
_____________________________________________________________________

2. Why salty DNA precipitates in alcohol whereas non-salty DNA does not?
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________

3. What can be the mechanism of breaking down cell membranes by dish soap?
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________

4. Why pineapple juice/papaya juice are added to the mixture? What are their roles in the DNA
extraction experiment?
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________

5. What did the DNA look like? Relate what you know about the chemical structure of DNA to
what you observed today.
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________
______________________________________________________________________________

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 Catanduanes State University
COLLEGE OF SCIENCE
Natural Science Department
Virac, Catanduanes

Conclusion:

Reference/s:

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