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CREATED BY:
CHEMICAL EDUCATION
TEACHER TRAINING AND EDUCATION FACULTY
SEBELAS MARET UNIVERSITY
SURAKARTA
2011
ISOLATION OF DNA FROM STRAWBERRIES
A. OBJECTIVES
Students knowing how to extract DNA
Student can see the precipitation process of DNA
Student can isolate DNA from strawberries
B. BASIC THEORY
Since DNA is the blueprint for life, everything living contains DNA. 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 the
field of biotechnology and forensics. Extraction and purification of DNA are the first
steps in the analysis and manipulation of DNA that 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 (sodiumdodecyl sulfate) is added. These solutions
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.
Strawberries, bacteria, humansall living things have genes, and all of these
genes are made of DNA. That's why scientists can take a gene from one living thing
and put it into another. For example, they can put human genes into bacteria to make
new medicines.
1 Releable bag 1
2 Test tube 4
3 Beaker glass 1
4 Meassuring glass 1
6 Pipette 4
7 Glass funnel 2
8 Filter paper 4
9 Stirer 2
2. Materials
Num
Name of Material Amount
b
1 Strawberries 1
2 Liquid soap 50 drops
3 Salt Sufficient
4 Water 5 ml
5 Ethanol 90% sufficient
D. PROCEDURE
Num
Step Image
b
7 Spool
E. OBSERVATION DATA
Num
Step Data
b
Place strawberries in a plastic Stroberries has broken become a little
1
resealable bag and smunch pieces.
Add liquid soap for amount : 5 There are a mixture between
2 drops, 10 drops, 15 drops, and 20 strawberry and soap
drops.
3 Add a small amount of water (5 ml) The mixture of strawberry is red
4 Add a pinch of salt Salt has disolved
Filter the strawberry mess Filtrate is red
5
The sediment has left in filter paper
Add 90% ethanol The mixture become two parts:
Top layer is clear solution and there
6
are white fiber in a mixture
Down layer is red
The DNA looked like white, thin
7 Spool fibers wadded up together forming a
clump.
F. DISCUSSION
This experiment has purposes to know how to extract DNA, to see the
precipitation process of DNA, and the main purpose is isolated DNA from strawberry.
This is a simple, effective protocol for spooling DNA. Ripe strawberries are an
excellent source for extracting DNA because they are easy to pulverize and contain
enzymes called pectinases and cellulases that help to break down cell walls. And most
important, strawberries have eight copies of each chromosome (they are octoploid), so
there is a lot of DNA to isolate.
The first step in this procedure is placed strawberries in a plastic and smunch.
This step has done to break the strawberry. It will destroy the cell wall and cell
membrane, which is used here is water and wrung. We recall the principles of cell
biology, namely diffusion-osmosis, where much less pressure coming in higher cells,
the cell will be damaged. In this step, supernatant / fluid contains organelles within the
cell and of course there is also DNA. But here the DNA was still inside the nucleus.
Then the second step is adding liquid soap for amount : 5 drops, 10 drops, 15
drops, and 20 drops. We know that DNA is still in the nucleus. Well, to destroy and
remove the nucleus of our DNA with the provision of soap treatment. This is because
most of the nuclear membrane is fat, to eliminate the fat we need soap. The soap helps
to dissolve the phospholipid bilayers of the cell membrane and organelles.
The next steps is adding a small amount of water and then add a pinch of salt.
The salt helps keep the proteins in the extract layer so they arent precipitated with the
DNA.
Then filter the strawberry mess. In this step we get the red filtrate while the
sediment or dregs left in the filter paper. After that add the 90% ethanol to the filtrate.
This addition resulted in the formation of two layers where the bottom layer is red
while the upper layer of clear liquid and there are white clumps of thread. Alcohol is
less dense than water, so it floats on top forming two separate layers. All of the grease
and the protein that we broke up in the first two steps move to the bottom, watery
layer. DNA will rise into the alcohol layer from the pea layer. DNA is not soluble in
ethanol. When molecules are soluble, they are dispersed in the solution and are
therefore not visible. When molecules are insoluble, they clump together and become
visible. The colder the ethanol, the less soluble the DNA will be in it yielding more
visible clumping. This is why it is important for the ethanol to be kept in a freezer
or ice bath.
The last step in this experiment is spooling. This step has purpose to collect the
DNA that has been isolated by all the procedure.
Single molecules of DNA are long and stringy. Each cell of the body contains
six feet of DNA, but it's only one-millionth of an inch wide. To fit all of this DNA into
the cells, it needs to be packed efficiently. To solve this problem, DNA twists tightly
and clumps together inside cells. Even when DNA extracted from cells, it still clumps
together, though not as much as it would inside the cell.
The DNA that occur is white, stringy stuff actually a mix of DNA and RNA.
That because of the procedure for DNA extraction is really a procedure for nucleic
acid extraction. However, much of the RNA is cut by ribonucleases (enzymes that cut
RNA) that are released when the cells are broken open.
G. ANSWER OF QUESTION
1. Place 1 strawberry in a plastic, resealable bag and smunch to break the strawberry
in order to destroy the cell wall and cell membrane.
2. Adding liquid soap has purpose to destroy and remove the nucleus of DNA with
the provision of soap treatment. The soap helps to dissolve the phospholipid
bilayers of the cell membrane and organelles.
3. Adding a small amount of water to help destroying the cell wall and cell
membrane of strawberry.
4. Adding a pinch of salt has to remove proteins that are bound to the DNA. It also
helps to keep the proteins dissolved in the aqueous layer so they dont precipitate
in the alcohol along with the DNA.
5. Filtering strawberry mess is to remove the larger particles from the solution, such
as seeds, pith, etc., allowing only the smaller cell components such as the DNA,
proteins, etc. to filter through.
6. Adding 90% alcohol causes the DNA to precipitate.
7. Spooling has done to take the DNA of strawberry that has been obtained.
H. CONCLUSION
1. DNA is is a nucleic acid that contains the genetic instructions used in the
development and functioning of all known living organisms. DNA is a large
polymer with unique physical characteristics of property.
2. Isolation of DNA can be done by simple method by using water, soap, salt, and
alcohol.
3. The function of adding some materials:
Soap, dissolve the phospholipid bilayers of the cell membrane and organelles
Water, help destroying the cell wall and cell membrane of strawberry
Salt (Sodium Chloride), remove proteins that are bound to the DNA
90% ethanol, causes the DNA to precipitate
4. In this experiment we get the DNA looked like white, thin fibers wadded up
together forming a clump.
I. BIBLIOGRAPHY
Aldina Wildan.2011. Lab report the basics of biotechnology. Semarang State
University
2010. DNA Isolation from Strawberry. http://www.science-projects.com/index.html
2011. DNA. http://en.wikipedia.org/wiki/DNA
Practicants