SPEAKER EUGENIO PEREZ NATIONAL AGRICULTURAL SCHOOL

Roxas, San Carlos City, 2420 Pangasinan, Philippines

Telephone No. : +6375•955•5258

Chapter I

Introduction

OBJECTIVE: To extract Deoxyribonucleic Acid (DNA) tissues of a living organism

(tomato).

Deoxyribonucleic Acid (DNA) is genetic material of all cellular organisms and

most viruses. DNA carries the information needed to direct protein synthesis and

replication. Protein synthesis is the production of the proteins needed by the cell or virus

for its activities and development. Replication is the process by which DNA copies itself

for each descendant cell or virus, passing on the information needed for protein synthesis.

In most cellular organisms, DNA is organized on chromosomes located in the nucleus of

the cell.A molecule of DNA consists of two chains, strands composed of a large number

of chemical compounds, called nucleotides, linked together to form a chain. These chains

are arranged like a ladder that has been twisted into the shape of a winding staircase,

called a double helix. Each nucleotide consists of three units: a sugar molecule called

deoxyribose, a phosphate group, and one of four different nitrogen-containing

compounds called bases. The four bases are adenine (A), guanine (G), thymine (T), and

cytosine (C). The deoxyribose molecule occupies the center position in the nucleotide,

flanked by a phosphate group on one side and a base on the other. The phosphate group

of each nucleotide is also linked to the deoxyribose of the adjacent nucleotide in the

chain. These linked deoxyribose-phosphate subunits form the parallel side rails of the

ladder. The bases face inward toward each other, forming the rungs of the ladder.
SPEAKER EUGENIO PEREZ NATIONAL AGRICULTURAL SCHOOL
Roxas, San Carlos City, 2420 Pangasinan, Philippines
Telephone No. : +6375•955•5258

The nucleotides in one DNA strand have a specific association with the

corresponding nucleotides in the other DNA strand. Because of the chemical affinity of

the bases, nucleotides containing adenine are always paired with nucleotides containing

thymine, and nucleotides containing cytosine are always paired with nucleotides

containing guanine. The complementary bases are joined to each other by weak chemical

bonds called hydrogen bonds. The genetic code in DNA is in triplets such as ATG. The

base sequence of that triplet in the partner strand is therefore TAC.Each strand consists of

alternating phosphate (PO4) andpentose sugar (2-deoxyribose), and attached on the sugar

is anitrogenous base. The DNA is found in thecytoplasm of prokaryotic cells, and chiefly

in the nucleus of eukaryoticcells. A small fraction of total DNA is present in

mitochondria andchloroplasts.DNA is a long polymer ofnucleotides to code for

thesequence of amino acid duringprotein synthesis. DNA is said to carry the genetic

‘blueprint’ since it contains the instructions or information (called genes) needed to

construct cellular components like proteins and RNA molecules.

SPEAKER EUGENIO PEREZ NATIONAL AGRICULTURAL SCHOOL
Roxas, San Carlos City, 2420 Pangasinan, Philippines
Telephone No. : +6375•955•5258

Chapter II

METHODOLOGY

This chapter discusses the tools and methods used in the experiment in obtaining the

results of the experiment.

Materials and Procedures

Tomatoes Blender Stirring rod

Pineale juice Chopping board Test tube

Salt Knife Test tube rack

Isopropyl alcohol Beaker Apron

Dish washing liquid Strainer Gloves

Dropper

Procedure

Three main procedures were employed in this experiment: (1) Lysing the cells, (2)

separating the DNA from other cell components, and (3) isolating the DNA.

A 100 mL of sliced tomatoes as DNA source, a 1 mL salt, and a 400 mL of tap water

were put together in a blender. The mixture was blended for 15 seconds, and then it was

strained in the beaker. Next, a 30 mL of liquid detergent was added in to the mixture. The

mixture was spun gently by the students and was let sat for 5 minutes. Afterwards, the

mixture was poured in to two test tubes evenly about one third fulls each and was added
SPEAKER EUGENIO PEREZ NATIONAL AGRICULTURAL SCHOOL
Roxas, San Carlos City, 2420 Pangasinan, Philippines
Telephone No. : +6375•955•5258

four drops of pineapple juice as the enzyme. The test tubes were tilted and were added

two drops of alcohol using a dropper until it had the same amount as the mixture.
SPEAKER EUGENIO PEREZ NATIONAL AGRICULTURAL SCHOOL
Roxas, San Carlos City, 2420 Pangasinan, Philippines
Telephone No. : +6375•955•5258

Chapter III

Results and Discussion

Deoxyribonucleic acid is further arranged by twisting and coiling upon itself with

various proteins associated to keep the strands organized and untangled. In its native

state, the sugar-phosphate backbone is the portion of DNA which is most closely exposed

to the environment, which is primarily water; in which DNA is soluble. Because of the

water’s overall polarity, the DNA is soluble in it. According to Paul Zumbo, all nucleic

acids are polar. In the case of DNA, the highly polar phosphate groups on the backbone

carry negative charges. This property accounts for water solubility, as water is also polar.

The positive charges of water interact with the negative charges of DNA and make a

solution. The DNA must be precipitated out of a solution with water to recover DNA for

further testing or visualization. This is accomplished by providing a stronger positively

charged in the solution since water has a relatively weak positive charge. Putting a salty

substance (Sodium) like salt will help stabilize the DNA from the nucleus.

The fact that the external membrane of cells and that of their nuclei are composed

of fatty substances (lipid molecules), the students used a simple detergent, particularly a

dishwashing liquid. A common detergent used in laboratories is the Sodium dodecyl

sulfate (SDS), but just a simple detergent will do (if you’re going to perform simple

extractions). In additional, students used enzyme because the cell is derived from a plant

material to digest the cell proteins and aid in precipitation.

SPEAKER EUGENIO PEREZ NATIONAL AGRICULTURAL SCHOOL
Roxas, San Carlos City, 2420 Pangasinan, Philippines
Telephone No. : +6375•955•5258

Once the DNA has been allowed to mix with water because it has been removed

from the nucleus of the cell, the introduction of sodium ions create a temporary attraction

between sodium and the backbone. The DNA is temporarily neutralized and then easily

disassociated from the water. At this stage, the introduction of an alcohol forces the DNA

and sodium ions to become even more tightly bonded, since alcohol is very non polar.

Isopropyl alcohol is what the students used, although ethanol can also be a candidate for

this. Finally, the DNA is precipitated in alcohol where it becomes visible.

The DNA was described as a long, stringy molecule, the clumps visible to the

glass are actually the tangled DNA molecules since salt was added to the mixture.
SPEAKER EUGENIO PEREZ NATIONAL AGRICULTURAL SCHOOL
Roxas, San Carlos City, 2420 Pangasinan, Philippines
Telephone No. : +6375•955•5258

Chapter IV

Conclusion and Recommendation

Like water, the DNA is a polar substance so it would not be visible. The best way

to visualize the DNA is to add a non-polar substance (ex. an isopropyl alcohol) and to

have an adequate amount of water. Moreover, then students should have worn gloves and

apron while performing the extraction since it’s the basic apparatuses that one should

have.

Chapter V

References

Redmond WA: Microsoft Corporation. (2008). Deoxyribonucleic Acid. Retrieved from

Microsoft Encarta Premium 2009.
Deoxyribonucleic Acid. (2018, January 8). Retrieved from www.biology-online.org
Deoxyribonucleic Acid definition. (2016, September 6). Retrieved from
www.medicinenet.com
How to Extract DNA. (2015). Retrieved from www.learn.genetics.utah.edu
How to Extract DNA from Fruits. (2007, January). Retrieved from www.funsci.com
Sodium Used in DNA extraction. (2016). Retrieved from www.sciencing.com
SPEAKER EUGENIO PEREZ NATIONAL AGRICULTURAL SCHOOL
Roxas, San Carlos City, 2420 Pangasinan, Philippines
Telephone No. : +6375•955•5258

The Deoxyribonucleic Acid (DNA) Extraction

Jacqueline Amor

Baron Gabuyan
SPEAKER EUGENIO PEREZ NATIONAL AGRICULTURAL SCHOOL
Roxas, San Carlos City, 2420 Pangasinan, Philippines
Telephone No. : +6375•955•5258

The Result
SPEAKER EUGENIO PEREZ NATIONAL AGRICULTURAL SCHOOL
Roxas, San Carlos City, 2420 Pangasinan, Philippines
Telephone No. : +6375•955•5258

Appendix A

Extraction of DNA

The sliced tomatoes as our DNA source Putting the sliced tomatoes, salt and water

The blending of mixture Straining the mixture to be putted in the beaker

SPEAKER EUGENIO PEREZ NATIONAL AGRICULTURAL SCHOOL
Roxas, San Carlos City, 2420 Pangasinan, Philippines
Telephone No. : +6375•955•5258

Adding the liquid detergent The mixture was spun gently

Adding 4 drops of pineapple juice Putting the same amount of alcohol as the mixture