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1.

0 Abstract

All cells in living organisms contain owe DNA that carrier information. In
this experiment we want to study DNA extraction from the fruits to prove
that there has DNA in eukaryote cells. We use orange as a sample during
experiment because it DNA is easy to conduct. First, before start
experiment, fill a cup of alcohol then put into the freezer about 30
minutes. After that, peel the orange into small pieces. Then, put the
orange the small pieces into the blender with warm water. Salt and
detergent are adding into the beaker of orange. Salt is used to neutralize
the negative charges on the DNA. In other hand, we also to study how
DNA works with present the three ingredients that mention early. Soap is
adding into the beaker to help the cell membrane broken apart and it
released DNA from cell. After 30 minutes, alcohol is adding into the orange
mixture with slowly pour to make the layer from orange mixture. As a
result, the DNA inside the fruits can be seen after we put on alcohol into it
mixture.

2.0 Introduction

DNA or called as deoxyribonucleic acid is located in living cell of


organisms. DNA also known as the blueprint for life because of the strand
is lies on it. Every protein is directly synthesis with DNA molecules and
contains all the genetics information in the new cell. In complex
eukaryotes such plants or fruits there are some DNA inside it. DNA does
not float around inside the nucleus of a cell. DNA is associated with a
variety of different proteins and encased in cellular membrane. DNA is an
incredibly small molecules but large in quantities that can be seen. The
objective in this experiment is to extract the DNA from fruits such as
orange, banana and apple.

DNA extraction is the process when the deoxyribonucleic acid (DNA) is


removed from the cells. In this experiment it divided by three basic steps
that in order to extract DNA from the fruits. We used extraction of DNA.
Extraction of DNA is the starting step in many diagnostic processes that

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used to detect bacteria in the fruit also environment as well as diagnosing
disease and genetic disorder. Firstly, before the experiment start fill a cup
of alcohol and put in freezer it about 30 minutes. Then, the cell must be
lysed (broken open) to release nucleus. Lastly, it precipitated out of
solution once DNA is release from fruits. In order to extract DNA from the
cell, the associated membranes and proteins must be removed from the
cell then physically separates from DNA so that it required the several
reagents as example salt, detergent and alcohol to complete the
extraction of fruits.

3.0 Objectives

1) To understand DNA is in almost all cells of living organisms including


the food.
2) To identify how each the ingredient that used in experiment may
help the DNA extraction process.
3) To extract DNA from the fruits.

4.0 Theory

DNA or deoxyribonucleic acid is the present in all cells of an organism that


basically hereditary material and it provides a blueprint for the cells
growth, function death and reproduction. DNAs structure is the double-
stranded helical structure that stated by Watson and Crick in 1953 (Dr
Ananya Mandal, 2012). At the Rockefeller Institute in 1929 by Phoebus
Levene come out with components that make up a DNA (Garbutt, 2012).
Firstly, components of the four bases which are Adenine (A), Cytosine (C),
Guanine (G), and Thymine (T), followed by sugar and last component is
phosphate. Each of the component unit is a nucleotide and the DNA
molecule consisted of string of nucleotide units that linked together
through the phosphate groups and these will form a backbone of the
molecule (Garbutt, 2012).

However, there are two scientists come out about DNA, Watson and Crick
in 1953, a model is created that showed a double helix with little rungs
and the two strands is connecting. The rungs were the bases of a

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nucleotide. They also stated that the paired of Thymine (T) with Adenine
(A) and Guanine (G) with Cytosine (C) DNA would look uniform. A
hydrogen bond is found that formed between two pairs of bases. In other
word, each side is a complement of the other (Garbutt, 2012).

In this experiment there are three ingredients that will give the impact of
DNA. We use detergent, salt and alcohol. The use of salt or sodium (NaCl)
is to neutralize the negative charges on the DNA. So the DNA strands
enables to stick together. That causes carbohydrates and proteins to
precipitate. Detergent in the soap disrupts the cells and nuclear
membranes of each cell to release the DNA. It does this by resolving lipids
and proteins that hold the membranes together.

5.0 Apparatus

Materials Apparatus
Orange Blender
Detergent (soap) beaker
Sodium chloride (salt) knife
Alcohol Warm water
Spatula
Glass rod

6.0 Procedures

DNA from fruits

1) Before start the experiment makes sure a cup of alcohol is filled and
placed it in the freezer about 30 minutes.
2) The orange or any fruits is peel off and cut into small pieces.
3) Small pieces of orange are put into a blender with small amount of
warm water. a teaspoon of salt is added into a blender then blend it
until a thick and easily pourable liquid is formed.
4) Over a glass jar is filtered the coffee position and the mixture is
poured into the filter until the jar fills about halfway.

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5) Two teaspoon of liquid dish soap is added into the jar and stirred it
carefully. During the stirring avoid from making bubbles.
6) After 30 minutes, the cold alcohol is removed from the freezer. Pour
it slowly into the orange mixture and down the side of the jar. For
this, an eye dropper is recommended to use.
7) Only used enough alcohol to form thin and separate layer on top of
the orange mixture
8) The mixture is left for 10 minutes and the mixture is undisturbed.
9) The DNA is coalesce and a long white strand is formed that float to
the top of the alcohol. A toothpick is used to pick it up for discuss in
report.

7.0 Result

Figure 7.1: white fluffy of DNA of orange.

8.0 Discussion
From the figure 7.1 shown that the white fluffy cloud, the present of the
DNA strands of the orange. Unfortunately, the picture that we was taken
look blurry so that the closely look DNA of orange is not really clear but in
the real we can see the DNA look like. The picture is blurry maybe
because of the angle that we take the picture and also the brightness in
the laboratory room is not good. The sample that we used in this
experiment is the orange. We used orange because large amount of DNA
inside the orange. So that it make it easier to extract DNA from it.

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In this experiment we required sodium chloride, NaCl added into the
mixture of orange after we blended the pieces of the orange. The purpose
we use the sample is to get the DNA that in in use into the DNA extraction
process which involve a series of the steps to gently break the cell, break
open the nuclear membrane, the DNA is separated from proteins and then
precipitate of the solution came out from it. So the used of sodium
chloride to stabilized the DNA after it has been stripped of its proteins and
aid in precipitation that is accomplished using the many chemical, based
on the structure of membranes, DNA and its electronegativity (Man,
2012). When the DNA has been removed from the nucleus of the cell and
mixed with water, sodium ions creates a temporary attraction between
sodium and the backbone. The DNA is neutralized and it disassociated
from the water with easily. The present of alcohol in the mixture of orange
forces the DNA and sodium ions to become more tightly bonded because
of very nonpolar alcohol (Taylor, 2010). In alcohol it could be present
ethanol or isopropyl alcohol depends on what we used in the laboratory.
The precipitate is out from the solution where it can be either
concentrated for purification or visualized by gently spooling it round a
smooth a glass rod once DNA is disassociated from the water and tightly
bound to the sodium.
The statement soap help break down the cell walls and membranes by
releasing DNA and other cell contents into the solution shown that
detergent in a soap will help the cell membrane broken apart and it
release the DNA from the cell. Detergent is the key ingredient that
dissolves the cell membrane so that the contents can escape. Detergents
have unique properties that enable manipulation of hydrophobic-
hydrophilic interactions among molecules. Detergent are amphipathic
molecules that means it contains a polar head and a nonpolar tail that
having aromatic character. The polar head group of ionic character form
the basis for broad classification of detergent for example it may be ionic,
non-ionic or zwitterions that having both positively and negatively charges
groups but that must be zero of a net charge (Detergent for The Cells Lysis
and Protein Extraction , 2016). When both soap and grease molecules

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organize themselves in the bubble with the head which like water and tail
which hate water so that their similar structures cause to combine and it
forming a greasy soapy ball when become more close to grease (Why Am I
Adding Detergent?, 2016). Then, it dissolves lipids with protein through it.

9.0 Conclusion
In conclusion, the objective in this experiment which to extract DNA from
the cells of orange is achieved. We had learnt a great knowledge in
extraction DNA. We also gained good knowledge to understand the
procedure and technique that required in this experiment. We observed
the result and study the results it. As a result in the figure 7.1 shown that
the picture if the white fluffy cloud that contains DNA strands. We also
able understanding the purpose we used detergents and salt as
ingredients that added in the mixture of orange. After we studied the all
ingredients, it is importance to success in the extraction process because
there is have their own objective when we adding detergent, salt and also
alcohol. We also get basic procedure of DNA that has made from Watson
and Cricks observation during their experiment in 1953.

10.0 Recommendations

1) Wear glove when handling ethanol to avoid skin contact with


ethanol because ethanol is the highly flammable and harmful
through skin contact.
2) Make sure ethanol has been cold first before use it because it will
protect DNA at low temperature by slowing down the activity of the
enzymes that may break it. It also helps DNA to precipitate quickly
in advance the most suitable temperature that to ensure the ethanol
will work is below 4 C
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3) The detergent that use in this experiment must be sit at least in 5
minutes to disrupt the cell membrane and nucleus membranes
because time is the once of the effect in order to complete the
extraction processes.
4) When DNA cant see by eyes clearly, stick or rod into the surface of
the soup and gently move it upward into the ethanol layer and

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make sure there are no bubble occurs because DNA are loosely
attached to the bubble so leave the mixture for 30 minutes that can
see more DNA precipitate.

11.0 References

1) Detergent for The Cells Lysis and Protein Extraction . (2016).


Retrieved MARCH 2016, from ThermoFisher SCIENTIFIC :
https://www.thermofisher.com/my/en/home/life-science/protein-
biology/protein-biology-learning-center/protein-biology-resource-
library/pierce-protein-methods/detergents-cell-lysis-protein-
extraction.html

2) Why Am I Adding Detergent? (2016). Retrieved March 2016, from


Learn.Genetics:
http://learn.genetics.utah.edu/content/labs/extraction/howto/deterge
nt/
3) Dr Ananya Mandal, A. (2012, 10 14). DNA Properties. Retrieved 3 21,
2016, from NEWS MEDICAL: http://www.news-medical.net/life-
sciences/DNA-Properties.aspx

4) Garbutt, C. (2012, 10 14). History of DNA Research. Retrieved 3 21,


2016, from NEWS MEDICAL : http://www.news-medical.net/life-
sciences/History-of-DNA-Research.aspx

5) Man, P. (2012, Sept 7). Work of Salt, Isopropanol and Ethanol in


Extarction DNA. Retrieved March 20, 2016, from G-BIOSCIENCES:
http://info.gbiosciences.com/blog/bid/156468/Work-of-Salt-
Isopropanol-and-Ethanol-in-DNA-Extraction

6) Taylor, S. (2010). Why is Sodium Used in Extraction DNA. Retrieved


3 20, 2016, from eHow :
http://www.ehow.com/about_6504902_sodium-used-dna-
extraction_.html

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12.0 Appendices

Figure 12.1: Smutch the orange using Figure 12.1: A cold


alcohol is added

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hand. into the
mixture of orange.

Figure 12.3: Detergent is added into the mixture of orange.

Figure 12.4: Orange is used as a sample