1

Pineapple Smoothie: DNA Isolation

Bergil Bernaldo, Bos Mauro Brandis, Jim Pauline Guiyab,
Krizelle Ann Hugo, Sheryll Ann Trinidad
Abstract
Deoxyribonucleic acid is the genetic material found in all cellular organisms. To find out
how the DNA looks like macroscopically, DNA isolation was performed on a pineapple fruit. The
isolation starts with breaking down of the fruit by turning it into smoothie, next is the addition of
ethanol in the pineapple puree destroys the lipids in the cells and separating the DNA. The
experiment had a control group using -naphtol, ammonium molybdate and phosphoric acid for
the positive control. The presence of DNA was determined by the change in color turning from
white to deep blue due to the reaction of the ammonium molybdate and -naphtol with the
phosphates acting as the backbone of the DNA. The experiment yielded a positive result
confirming the presence of DNA in the isolated spooled material.
Keywords
Deoxyribonucleic acid; Pineapple (Anasas comosus); nucleoproteins; -naphtol; Ammonium
molybdate;
Introduction
All known cellular life and some viruses contain DNA (deoxyribonucleic acid). DNA is a
nucleic acid that contains the genetic instructions for the development and function of living
things.
[1]
It has a double helix structure and is made up of chemical building blocks called
nucleotides. These building blocks are made of three parts: a phosphate group, a sugar group and
one of four types of nitrogen bases.
[2]
The four types of nitrogenous bases are; adenine (A),
guanine (G), thymine (T), and cytosine (C).
The Biochemistry students of BS Chemistry 3-1 were asked to isolate DNA from pineapple
(Ananas comosus) fruit. A pineapple fruit is actually multiple fruits that have been formed from a
thickened stem surrounded by hundreds of fruitlets that have been fused together. Pineapples
have a tough and hard rind, which consists of the remains of the flowers and the pointed bracts.
The color is golden orange and, despite its hard and dry appearance, juice can be extracted from
the rind. Pineapple has no seeds but the core of the fruit is hard since it consists of the old stem.
Removal of the core produces the characteristic pineapple rings. The flesh is juicy and yellow.
[3]
2
DNA isolation is the process of extracting DNA from a cell. It involves separating DNA
from other cellular components. DNA is isolated by placing the cells in a tube containing a
special solution and mechanically or chemically breaking the cells open. This causes the cell to
release its contents into the solution which may contain enzymes, chemicals, and salts. Enzymes
are used to chew up the proteins; chemicals to destroy any RNA present and salts to help pull the
DNA out of solution. The DNA will exist in long strands that form a mucous-like glob.
[4]
A phosphate test using ammonium molybdate and -naphthol can be used to confirm the
presence of DNA.
Methods
Add 3-5 drops of Nitric Acid
then apply constant heat.
A
Collect the ethanol that contains the precipitate
and place in a separate test tube.
Run the sample in the centrifuge for 2
minutes.
Dispose the ethanol to isolate the
spooled material.
Add 3-5 drops of Sulfuric Acid.
Put the sample in a test tube. It
should occupy 1/3 of the test
tube.
START
Prepare chunks of pineapple then
puree.
Filter the puree by using cheesecloth
then collect the filtrate
Add liquid detergent to
destroy the lipids.
Add ice cold 95% Ethanol.
Submerge the sample in a water
pan that contains ice.
Let the sample sit for a couple of
minutes.
Do you observe
white precipitate in
the ethanol?
No
A
No
Does the color of the
sample become clear pale
yellow?
Yes
No
A
Add distilled water and some drops of
Ammonium Molybdate.
Does the color of the
sample become clear?
Yes
3
End
A
a
Add -napthol
Apply heat until -napthol
dissolves.
Compare the result with the control
set-up.
C
a
B
a
Start
Put 1ml of distilled water in a test tube.
Add 1ml of Ammonium
Molybdate and a small piece
of -napthol.
Heat until -napthol dissolves.
B
a
Start
Put 1ml of Phosphoric Acid in a test
tube.
Add 1ml of Ammonium
Molybdate and a small piece of
-napthol.
Heat until -napthol dissolves.
C
a
Add water to dilute
4
Results and Discussions
A. Liberation of Nucleoproteins
For approximately two minutes, white spool-fiber known as nucleoproteins precipitated
in the ethanol layer of the test tube. The nucleoproteins contain the DNA material needed in
the following test.
B. Confirmation for presence of DNA
The clear color of the nucleoprotein aliquot remained clear after few drops of sulfuric
acid was added. Upon heating the test tube, drops of nitric acid was added until yellow fumes
disappeared turning the color of the solution into light yellow. The color of the solution
turned clear upon heating it with few drops of ammonium molybdate. The solution turned
dark blue when heated after a small amount of -naphtol was added. The solution was diluted
to visualize better the blue color that indicates a positive result. The presence of the DNA is
confirmed because of the phosphate backbone in the DNA that reacted with the reagents.
References
[1] DNA. http://www.sciencedaily.com/articles/d/dna.htm (accessed December 7, 2013)
[2] Deoxyribonucleic Acid (DNA) Fact Sheet. http://www.genome.gov/25520880 (accessed
December 7, 2013)
[3] Characteristics of a Pineapple | eHow. http://www.ehow.com/info_8553867_characteristics-
pineapple.html (accessed December 8, 2013)
[4] http://oklahoma4h.okstate.edu/aitc/lessons/intermed/DNA_Blueprint.pdf
Figure 1: Blue color of the positive control
(left) and blue color of the positive result
(right).
Figure 2: Addition of nitric acid in acid digestion.