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08
Isolation and Chemical Characterization of DNA
I. Objectives
II. Introduction
The most characteristic constituents of the nucleus are the nucleic acids, DNA and
RNA. Their function have always been of great interest, because of the presence of DNA in
the Chromosomes, and in the transmission of hereditary characteristics. DNA Isolation is one
of the most basic and essential techniques in the study of DNA. The extraction of DNA from
cells and its purification are of primary importance to field of biotechnology and forensics.
Extraction and purification of DNA are the first steps in the analysis and manipulation of
DNA they allow scientists to detect genetic disorders, produce DNA fingerprints of
individuals, and even create genetically engineered organisms that can produce beneficial
products such as insulin, antibiotics and hormones.
DNA can be extracted from many types of cells. The first step is to lyse or break open
the cell. This can be done by grinding a piece of tissue in a blender. After the cells have
broken open, a salt solution such as NaCl and a detergent solution containing the compound
SDS (Sodium dodecyl Sulfate) is added. These solution break down and emulsify the fat &
proteins that make up a cell membrane. Finally, Ethanol is added because DNA is soluble in
water. The Alcohol causes DNA to precipitate, or settle out of the solution, leaving behind all
the cellular components that aren’t soluble in alcohol. The DNA can be spooled (wound) on a
stirring rod and pulled from the solution at this point.
III. Materials
A. Equipment
B. Reagents
IV. Procedure
A. Extraction of DNA
1. Hydrolysis
V. Results
The precipitation of DNA was due to the addition of ice-cold ethanol to the solution. DNA
have an ionic nature which makes it insoluble in an aqueous medium which was made less polar by
addition of an organic solvent. Therefore, a white thread-like precipitate was formed. deoxyribose
and it was same with what happened to the standard solution, which indicated a positive result.
Meanwhile, in the test for purines, the DNA color turned from brown to colourless while the
standard guanine solution turned from purple to brown residue which shows a negative result. In
the test for pyrimidine, both the standards formed a purple precipitate while the DNA formed a
white precipitate in a colourless solution which indicates a negative result. The onion DNA showed a
positive result in the test for deoxyribose because the sugar found in the DNA is D-deoxyribose
which reacts with the diphenylamine reagent. However, the visual result should be a blue colored
solution. On the other hand, the DNA must have a positive result with the test for purines and
pyrimidines because the DNA are composed of four bases which are guanine, thymine, adenine and
cytosine.
VII. Conclusion: DNA reacts in acid hydrolysis and less reactive in basic hydrolysis since it is
negatively charged because of the phosphates surrounding it, which means that its isoelectric pH is
above 7 making it basic. DNA is ionic in nature making it insoluble in less polar aqueous medium. As
a result, DNA precipitated out in the presence of ice-cold ethanol because all the other components
of the mixture stayed in the solution except for DNA. The erroneous color results of the standards in
the chemical characterization was because of the reagents used. They might be contaminated or
were unused for a very long time. Because theoretically, a blue solution must be seen as a positive
result in dische test, a red residue in the murexide test, and a purple coloration in the wheeler-
johnson test. Thymine cannot be determined through the test for pyrimidines since it has a methyl
group which prevent its interaction with excess of bromine water. Meanwhile, the error in the DNA
was because of the improper execution of the methods by the experimentalists.
5-carbon sugars ribose and deoxyribose are important components of nucleotides, and are found
DNA, respectively.
2.what is the purpose of each of the following components in this protocol? Dishwashing liquid, salt,
meat tenderizer (pineapple juice), ethanol.
Dishwashing liquid-removes fats plus protein to extract Dna meat tenderizer source of Dna Salt-
softens phospholipids in membrane Ethanol-separates Dna from other cell components shows Dna.
3.We can’t really see a Dna molecule under the microscope unless it is rightly coiled into a
chromosome. Why can you see the Dna After you put it into the ethanol?
Dna is released from the wheat ram mixed with water and detergent. The released Dna is
precipitated into the alcohol with after cell components.
4.if you were able to spool the Dna you could see that it is stringly and has the consistency of thick
syrup or mucus. Based on what you know about the molecule; why do you think it has this
consistency?
Dna has a double helix and is like a sparling string. It’s string because it’s wrapped around the
histones which mate s up the chromosomes.
XI. REFERENCES
oyer, R. (2009). Biochemistry Laboratory: Modern Theory and Techniques. San Francisco,
USA: Pearson Education, Inc.,.
Bruns, D., Burtis, C., Ashwood, E., & Sawyer, B. (2007). Fundamental of Molecular
Diagnostics. Philadelphia, PA: Saunders Elsevier.
Hecht, S. (1996). Bioorganic Chemistry: Nucleic Acids. New York, NY: Oxford University
Press.
X. Conformer