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Strawberry and Blueberry DNA Extraction

Lab Pt 1 - Strawberry DNA Extraction

Overview:
Every living thing is made of many many cells. Each cell has its own job and function
controlled by the nucleus or brain of the cell. More specifically, cells are controlled by
DNA (deoxyribonucleic acid) which is able to code every cell with a certain function.
Wound very tightly, long strands of DNA are called chromosomes. In cultivated
strawberries there are eight copies of genetic material making them an octoploids. In
humans we only have two sets making us diploids. As a result, when extracting DNA it is
easier to find strawberries DNA because there is so much of it.

Problem Statement:
Is it possible to extract DNA from a strawberry using the buffer solution and ethenol?

Hypothesis:
If the experiment is conducted correctly then it will be possible to extract DNA from the
strawberry because the buffer solution that is mixed with the crushed strawberry is made
from salt and soap which breaks down the cells in the strawberry and allows access to the
DNA contained in the nucleus.

Independent Variable:
● DNA buffer
● Ethenol
Dependant Variable:
● Observations
● Extracted DNA
Materials:
● heavy duty ziploc bag (freezer or storage bag)
● 1 strawberry
● DNA extraction buffer (900mL water, 50mL dishwashing detergent, 2 teaspoons
salt)
● beaker
cheesecloth to fit in small funnel (4” X 4” should be appropriate)
● small funnel
● 50mL vial / test tube
● glass stirring rod
● cold ethanol (isopropyl alcohol)

Procedure:
1. Place one strawberry in a Ziploc bag.
2. Smash/grind up the strawberry using your fist and fingers
for 2 minutes. Careful not to break the bag!!
3. Add the provided 10mL of extraction buffer (salt and
soap solution) to the bag.
4. Kneed/mush the strawberry in the bag again for 1 minute.
5. Assemble your filtration apparatus as shown to the right.
6. Pour the strawberry slurry into the filtration apparatus and let it drip directly into your
test tube.
7. Slowly pour cold ethanol into the tube. OBSERVE.
8. Dip the loop or glass rod into the tube where the strawberry extract and ethanol layers
come into contact with each other. OBSERVE.

Observations:

Lab Step Observations

Step 2: Grind up strawberries The Strawberry was easy to grind which


allowed for better access to the cells. It
created a mush the kind of looked like a
smoothie. Dark red becuase of the juice.

Step 3: Add solution When the buffer solution made contact


with the Strawberry it got foamy and
mixed in well. The more I mixed the more
bubbles appeared. Must be from the soap.
The whole solution turned a light pink
color because of the suds

Step 7: Pour Ethanol The ethanol immediately separated the


water and DNA allowing us to see it and
easily remove it from the tube. It looked
like all the juice from the strawberry
separated making it look like cranberry
juice with the DNA at the bottom.
Analysis:
DNA is impossible to see with the naked eye, even when pulverizing the strawberry.
However when wound together and in large quantities DNA strands become much more
obvious and are even touchable.

Supporting Evidence of Hypothesis:


The hypothesis states “If the experiment is conducted correctly then it will be possible to
extract DNA from the strawberry because the buffer solution that is mixed with the
crushed strawberry is made from salt and soap which breaks down the cells in the
strawberry and allows access to the DNA contained in the nucleus.” This hypothesis was
correct because based on research focusing on breaking down cells, and when the lab
concluded, there was extracted DNA visible to the naked eye and touchable.

How Does The Buffer Solution Work?


The buffer solution focuses on two parts: Breaking down the cell wall then isolating the
DNA. The first step of the the DNA extraction process is to break down the cells outer
shell and first layer of protection called the phospholipid bilayer. This bilayer is made up
of hydrophobic and hydrophilic lipids. When a cell is exposed to water it makes a layer
of lipids around the cell whose hydrophilic heads can be exposed to the water and the
hydrophobic tails can be tucked between the two layers.
Soap is also made of hydrophilic and phobic lipids. The difference between cell and soap
lipids this that soap lipids are much more soluble in water than cell lipids. However, soap
lipids also have hydrophobic tails. This similarity means that when soap and cell lipids
come in contact they can combine but the solubility of the soap lipids pulls the cell lipids
apart and into the water. This is what allows the cell wall to be broken down.
The next step in extracting DNA is being able to isolate it from the rest of the contents of
the cell. Unlike the interior of the cell and its parts, DNA is only soluble in fresh water
not salt water. This means that when in contact with the salt water in the buffer solution,
the DNA becomes free standing while the rest of the cell mixes into the water.

Major Findings:
● DNA is only soluble in fresh water
● Ethenol is to spereate DNA from water
● DNA becomes a precipitate in Ethenol
● Cell wall is called a phospholipid bilayer
● Soap breaks down cells
● DNA is small but in large number is visable and feelable.

Conclusion:
The DNA extraction lab is a great way to see and interact with DNA first hand as well
and understand how a cell works depending on the environment it is exposed to. In the
lab there were a few key parts that allowed for the lab to work. The First is the
pulverization of the strawberry. This step was crucial because since strawberries are
plants their cells have a rigid cell wall layer that would make it hard to break down the
cell. By smashing the strawberry, the rigid cell wall found in plants is broken and allows
for the cell to start being broken down even further. Next is adding the buffer solution
that helped isolate the DNA and break down the cell. Another important step is filtering
the strawberry-buffer solution. By filtering the solution we are able to keep any large un
broken down pieces of strawberry out of the final mixture as well as only let the smallest
things through like the DNA. Finally, it was crucial to add the ethanol to the mixture
because while DNA is soluble in water, it isn't in alcohol. This means that when the
ethanol is poured into the strawberry- buffer solution, the ethanol is able to pull the DNA
from the water and keeping it secluded. The ethanol is also able to slow down any
damaging reactions that may cause harm to the DNA. As a result it is easy to find and
pick out the DNA which should now look like whitish mucus.

DNA is constructed in a twisted ladder structure known as a double helix. When looking
at the DNA extracted from the strawberry it looks nothing like that. This is because there
is so much DNA all tangled together in one area. Yet, looking closer, is the DNA mucus
was pulled apart it and looked at under a microscope the double helix structure would
still be there for every strand of DNA. This can be thought of in terms of string. “A
person cannot see a single cotton thread 100 feet away, but if you wound thousands of
threads together into a rope, it would be visible much further away.” This is analogous to
DNA because as stated before, individual DNA is impossible to see with the naked eye,
but when wrapped together is exorberant amounts, DNA is quite easy to see. However
just like with the rope, It will still be impossible to see individual strands or pieces of
string. Only the final mass will be visible.

It is important for scientists to be able to remove DNA for analysis. In order to


understand and map genetic dispositions, scientists need to be able to actually see strands
of DNA and compare them to one another to see differences and anomalies. Another
important part is to see the effects of medical treatment and advancement found in DNA.
If DNA is a map of humans code then being able to read it and see the effects of different
treatment is important to helping improve and take care of humans.

Recommendations for Lab Improvement:


In order to improve the lab I would find a more effective way to mix the buffer and
stawberry solution. The reason for this being that when the buffer mixture was put witht
eh strawberry it was very sudsy and hard to stain through the cheesecloth as well as mix
in. Maybe a solution to this problem would be to make the buffer solution with the
strawberry mixed in, or another option would be some way to remove the suds. Other
than that the lab was helpful and effective.
Lab Pt 2 - Blueberry DNA Extraction

Overview:​ Many plants are polyploids meaning they have more than two sets of genetic
information. This is true in the case of the common blueberry also known as Vaccinium
angustifolium. This blueberry is a tetraploid meaning it has four copies of genetic
information. This excess DNA will make it easier to view when extracted from the
blueberry. However, since it has only half the amount of genetic information as the
strawberry there may be less of it.

Problem Statement:
Is it possible to extract DNA from a strawberry using the buffer solution and ethenol?

Hypothesis: ​If the lab is conducted correctly than blueberry DNA will be extracted
because based on the chemical components of how the procedure is able to release
strawberry DNA it should work on blueberries as well especially since they also have lots
of DNA.

Independant Variable:
● DNA Buffer
● Ethenol
Dependant Variable:
● Observations
● Extracted DNA
Materials:
● heavy duty ziploc bag (freezer or storage bag)
● 3 blueberries
● DNA extraction buffer (900mL water, 50mL dishwashing detergent, 2 teaspoons
salt)
● beaker
cheesecloth to fit in small funnel (4” X 4” should be appropriate)
● small funnel
● 50mL vial / test tube
● glass stirring rod
● cold ethanol (isopropyl alcohol)

Procedure:
1. Place all blueberries in a Ziploc bag.
2. Smash/grind up the blueberries using your fist and fingers
for 2 minutes. Careful not to break the bag!!
3. Add the provided 5mL of extraction buffer (salt and
soap solution) to the bag.
4. Kneed/mush the blueberries in the bag again for 1 minute.
5. Assemble your filtration apparatus as shown to the right.
6. Pour the blueberry slurry into the filtration apparatus and let it drip directly into your
test tube.
7. Slowly pour cold ethanol into the tube. OBSERVE.
8. Dip the loop or glass rod into the tube where the blueberry extract and ethanol layers
come into contact with each other. OBSERVE.

Observations:
Lab Step Observations

Step 2: Grind up blueberries The blueberry was easy to grind which


allowed for better access to the cells
however there was very little. Mostly all
there was was the leftover blueberry skins
which were filtered out

Step 3: Add solution When the buffer solution made contact


with the blueberry it got foamy and mixed
in well but with less blueberry I put in less
solution which was a good thing but a
very small amount over all

Step 7: Pour Ethanol The ethanol immediately separated the


water and DNA allowing us to see it and
easily remove it from the tube even if
there was very little of it. There wasn't
much color either.
Analysis:
DNA is impossible to see with the naked eye, even when pulverizing the blueberry.
However when wound together and in large quantities DNA strands become much more
obvious and are even touchable. The only problem with using three blueberries is that the
blueberries themselves are very small and therefore produced little DNA in comparison
to the strawberry.

Supporting Evidence of Hypothesis:


The hypothesis states “​ ​If the lab is conducted correctly than blueberry DNA will be
extracted because based on the chemical components of how the procedure is able to
release strawberry DNA it should work on blueberries as well especially since they also
have lots of DNA.” This hypothesis was correct because based on research focusing on
breaking down cells and when the lab concluded, there was extracted DNA visible to the
naked eye and touchable.

How Does The Buffer Solution Work?


The buffer solution focuses on two parts: Breaking down the cell wall then isolating the
DNA. The first step of the the DNA extraction process is to break down the cells outer
shell and first layer of protection called the phospholipid bilayer. This bilayer is made up
of hydrophobic and hydrophilic lipids. When a cell is exposed to water it makes a layer
of lipids around the cell whose hydrophilic heads can be exposed to the water and the
hydrophobic tails can be tucked between the two layers.
Soap is also made of hydrophilic and phobic lipids. The difference between cell and soap
lipids this that soap lipids are much more soluble in water than cell lipids. However, soap
lipids also have hydrophobic tails. This similarity means that when soap and cell lipids
come in contact they can combine but the solubility of the soap lipids pulls the cell lipids
apart and into the water. This is what allows the cell wall to be broken down.

The next step in extracting DNA is being able to isolate it from the rest of the contents of
the cell. Unlike the interior of the cell and its parts, DNA is only soluble in fresh water
not salt water. This means that when in contact with the salt water in the buffer solution,
the DNA becomes free standing while the rest of the cell mixes into the water.

Major Findings:
● DNA is only soluble in fresh water
● Ethenol is to spereate DNA from water
● DNA becomes a precipitate in Ethenol
● Cell wall is called a phospholipid bilayer
● Soap breaks down cells
● DNA is small but in large number is visable and feelable
Conclusion:
The lab was a success. Based on the trial run done on the stawberry, it seemed highly
likeley. Both with plant structures, normal cell functions, and based the reactions given
by the strawberry cells, the blueberry cells would behave the same way. The only
difference that was apparent was the amount of DNA in the end of the experiment. With
the strawberry there was a large amount of DNA and it was easy to pick up whereas the
blueberry DNA was harder to find and collect. After research I base this on two factors.
The first being that the total mass of the three blueberries was not nearly the mass of the
strawberry meaning that in total the blueberries had less cells and thus less DNA to
extract. The second is that blueberries are only tetraploids whereas strawberries are
octoploids. Whilst conducting the lab the thought occurred to me that if I were to be able
to somehow switch the DNA in the blueberries cells would they still function correctly?
The answer I came up with was that they would as long as there was a correct amount of
genetic information present. I also came to this answer because assuming the blueberries

all came from one parent plant, they should contain the same genetic information.

Recomendations for Lab Improvement:


The only thing I would change concerning the blueberry part of this lab would be the
amount of blueberries used. In my experiment I used three which worked but it was
harder to see the DNA clearly. I would use more to be able to have more DNA present to
extract and observe.
Works Cited

“DNA Extraction.” ​SpringerReference​, doi:10.1007/springerreference_66580.

“How Does Soap Affect Membrane Permeability? Which Component of the Membrane
Does It Affect?” ​Biology Stack Exchange​, 20 Oct. 2015,
biology.stackexchange.com/questions/39760/how-does-soap-affect-membrane-per
meability-which-component-of-the-membrane-does. Accessed 30 May 2017.

Ballington, James R. “Blueberry.” ​AccessScience​, doi:10.1036/1097-8542.088100.


Accessed 30 May 2017.

“Fruit Genetics Friday #1b: Polyploidy(And Yeah, I Know It's Saturday...).” ​The Fruit
Blog,​ Evil Fruit Lord,
thefruitblog.blogspot.com/2006/05/fruit-genetics-friday-1b-polyploidyand.html.
Accessed 30 May 2017.

Lyrene, P.M., et al. “Polyploidy and Sexual Polyploidization in the Genus Vaccinium.”
SpringerLink​, Kluwer Academic Publishers,
link.springer.com/article/10.1023/A:1025608408727. Accessed 30 May 2017.

Mweddin. “Putting the Pieces Together: Sequencing the Blueberry Genome.” ​Plants for
Human Health Institute,​ Slice of PHHI, 23 Sept. 2011,
plantsforhumanhealth.ncsu.edu/2011/09/23/sequencing-the-blueberry-genome/.
Accessed 30 May 2017.

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