Title DNA Extraction

Objective To extract the DNA from various plant samples.

Introduction All living things are made of tiny building blocks called cells. Each cell contains inherited genetic information, packaged in the form of genes. A gene is made of a length of DNA (deoxyribonucleic acid) that has a message encoded in its chemical structure. DNA is a double-stranded, helical nucleic acid molecule capable of replicating and determining the inherited structure of a cells protein (Boyer, 2006). DNA is the genetic material that organisms inherit from their parents. It contains information used in everyday metabolism and growth and influences most of our characteristics.

We are made up of billion cells. Most of the DNA located in the nucleus. Nucleus contains our unique set of 46 chromosomes. DNA is so tightly coiled that approximately 1.8 meters of it is able to fit into the nucleus of a human cell (DNA and Gene, 2008). DNA stores all the coded information needed for everyday growth and metabolism.

All the nitrogenous bases in DNA are derivatives of the two heterocyclic compounds purine and pyrimidine. The major purines in DNA are adenine (A) and guanine (G); the major pyrimidines in DNA are thymine (T) and cytosine (C) (Campbell, 2008). Wherever one strand of a DNA molecule has an A, the partner strand has a T. And a G in one strand is always paired with a C in the complementary strand. Therefore, in the DNA of any organism, the amount of adenine equals the amount of thymine, and the amount guanine equal the amount of cytosine.

Although the base pairing rules dictate the combinations of nitrogenous bases that form the rungs of the double helix, they do not restrict the sequence of nucleotides along

each DNA strand. The linear sequence of the four bases can be varied in countless ways, and each gene has a unique order, or base sequence. (Campbell, 2008)

The process of extracting DNA is to separate DNA from the unwanted substances of the cell gently enough so that the DNA is not broken up. This means that DNA extraction is the removal of deoxyribonucleic acid (DNA) from the cells or viruses in which it normally resides. Extraction of DNA is often an early step in many diagnostic processes used to detect bacteria and viruses in the environment as well as diagnosing disease and genetic disorders.(Rice, 2010).

Materials y y y y y y y Onion Detergent Salt solution Alcohol Test tubes Wooden stick Filter paper

Methods 1. A piece of onion in grinded up in small amount of water. 2. Some detergent and salt solution were added and stirred gently for 5 minutes. 3. The liquid was decanted from the mixture into a clean test tube (filter paper was used to get clear supernatant). 4. Cold alcohol was added in by tilting the test tube at a 45-degree angle and the alcohol was poured very slowly down the side of the tube. The alcohol should just trickle down the side and come to rest on the top of the water so that it forms a separate layer ( the layer not allow to mix) 5. The test tube was placed in its rack and do not move it for at least 15 minutes. The DNA will begin precipitating out immediately between the two layers of liquid. 6. After 15 minutes, the DNA was floated on the top of the test tube. A wooden stick was used to spool it cotton candy. 7. The DNA was described.

Result

Conclusion The DNA of onion was extracted. It appears as a white,stringy mass with small bubbles formed on it.

Questions 1. What does the DNA look like? DNA molecule is very long, it is stunningly simple. DNA looks like an incredibly long twisted ladder. This shape is called a double helix.The sides of the ladder are a linked chain of alternating sugar and phosphate molecules. The rungs connect to the sugar molecules and are known as bases.There are four bases - adenine (A), thymine (T), guanine (G) and cytosine (C). Each rung is made up of two bases that link together. Because of their chemical nature, A will only link with T and G will only link with C.

2. What do you think was the specific purpose of adding each of the following (i) detergent, (ii) salt, (iii) alcohol? I. Detergent The detergent causes the cell membrane to break down by dissolving the lipids and proteins of the cell and disrupting the bonds that hold the cell membrane together. The detergent then forms complexes with these lipids and proteins, causing them to precipitate out of solution.The detergent also inactivates enzymes called nucleases,that would destroy the DNA.DNA is soluble and more stable under slightly basic conditions.So the detergent solution is buffered to pH 8.

II. Salt Salt provides the DNA with a favorable environment, so that the DNA can extract easily. It contributes positively charged atoms that neutralize the normal negative charge of DNA. The salt shields the negative phosphate ends of DNA, which allows the ends to come closer so the DNA can precipitate out of a cold alcohol solution.

III. Alcohol The alcohol is used to extract the DNA from the onion juice. The reason of using alcohol is to stay on top of the onion juice is because by doing that the liquid will form two distinct layers. Generally, molecules are attracted to the boundaries of two liquids.DNA will not dissolve in this alcohol, so the DNA comes out of the solution,

or precipitates. It is less dense than water or cell scum which is what settles to the bottom of the test tube.So it floats up into the alcohol layer, where like a snotty, string-like substance, with small bubbles formed on it.

3. Why is onion suitable for DNA extraction? An onion is used because it has a low starch content, which allows the DNA to be seen clearly.

4. What is the component in the detergent that helps in the DNA extraction? The enzymes that contains in the detergents will helps in DNA extraction.Besides that, the hydrocarbon chain in the detergents will destroy and break the membrane then clump together. Detergents have hydrophobic or water-hating molecular chains and hydrophilic or water-loving components.As the head was hydrophilic, so they will face outside forming a circle like shape then as the water molecule pulling the polar head, the membrane together with the hydrocarbon chain will pulled outand the membrane burst.

Reference 1. Campbell,N.A.,Reece,J.B.,&Mitchell,L.G.(2008), Biology. (8th ed.), San Francisco: Pearson/Benjamin Cummings. 2. Rice, G. (2010, August 27). DNA Extraction. Retrieved August 1, 2010, from Microbial Life Educatinal Resources: http://serc.carleton.edu/microbelife/research_methods/genomics/dnaext.html 3. Boyer, R. (2006). Concepts In Biochemistry (3rd ed). John Wiley (Asia): Wiley Asia Student Edition 4. DNA and Gene. (2008, June 30). Retrieved August 3, 2011, from Biotechnology Online: http://www.biotechnologyonline.gov.au/biotechnologyonline/biotec/dnagene s.html