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BTEC Applied Science Unit 11 Assignment 4
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DNA ExtractionIntroduction:
DNA is a double helix structure that consists of long and interconnected chains of
nucleotides. Every living organism possesses a unique set and arrangement of nucleotides
and nitrogenous bases. DNA is able to be extracted from organisms by breaking down the
cell membrane and separating out the DNA structure from the other parts of the cell.
What is DNA Extraction? How is it used in Medicine?
DNA extraction is the process of removing the deoxyribonucleic acid from the cells of an
organism. DNA extraction is most often the earliest step in a range of diagnostic processes
that are used to detect pathogens or viruses in the host. It can also be used to diagnose
diseases and genetic disorders such as cystic fibrosis, Huntington’s or Down syndrome.
DNA extraction can also be used to identify if the patient is a carrier or sufferer.
Equipment
● Raspberries
● Boiling tube
● Glass rod
● Salt/detergent mix
● Blender
● Coffee filter paper
● Funnel
● Beaker
● Protease enzyme
● Ethanol
● Pipette
Method:
1. Put 5 raspberries in a beaker and smash them together to break - This is done to
increase the surface area
2. Add salt/detergent mix to the raspberries and mix again - This is done because it will
neutralise the charge of the DNA and make the molecules less soluble in water and
remove the proteins that are bound to the DNA and keep the proteins dissolved in
the beaker of water
3. Blend the raspberries in a blender - This exposes a greater surface area from which
we can extract DNA
4. Place a coffee filter in a funnel and pour the mixture into the funnel - Filtering the
mixture separates the liquid from the solids as we’re only interested in the filtered
liquid that contains the DNA
5. Add 5 drops of protease enzyme using a pipette to the liquid - This is so that it
degrades the DNA associated proteins
6. Add solution to boiling tube
7. Tilt the boiling tube gently at an angle and keep adding drops of ethanol until there is
a separating layer - DNA will precipitate at the boundary between the two layers
What is PCR? How is it used in Medicine
, In molecular biology, there is a simple yet useful procedure known as the polymerase chain
reaction or PCR for short. PCR is a process that allows us to amplify countless copies of
segments of DNA from a small sample. PCR imitates what happens in cells when DNA is
replicated before cell division. However, the PCR process is carried out in controlled
laboratory conditions. They use a PCR machine, or thermocycler, to put in boiling tubes that
contain the DNA mixture. The machine adjusts the temperature to suit each step of the
procedure.
○ PCR Process
Denaturation: First, much like DNA replication, the double helix structure of
the two strands must be unwound. This is done by increasing the temperature
of the mixture so that the hydrogen bonds between the complementary bases
strat to break.
Annealing: Second, the primers must bind to the target DNA sequences and
begin polymerisation. Then, one primer binds to each strand; however, all of
this can only occur once the temperature has been lowered.
Extension: Finally, using the original strands as templates, we can make new
strands of DNA. After that, a DNA polymerase enzyme binds the unoccupied
nucleotides together. The nucleotides go in the same order as the original
strand. In one cycle of PCR, the result is two double-stranded sequences of
target DNA where they contain one newly made strand and one original
strand. The PCR recycle is repeated usually 20-30 times as it only takes 2-3
hours to get a billion copies of DNA.
What is Transformation of Cells? How is it used in Medicine?
Bacterial cells are able to acquire foreign DNA in a process known as transformation.
Specifically prepared bacteria are mixed with human DNA. The bacteria are then given a
heat shock which allows an opening on the structure of the cell for the plasmids to enter
from.
Transformation of cells is primarily used for cloning DNA in order to produce large amounts
of specific human proteins. For example, those who suffer from diabetes are not able to
produce their own insulin. This means insulin needs to be injected into their bodies. The
medicine industry uses cell transformation to manufacture insulin and sell them on the
market for exaggerated prices.
○ DNA Cloning Process:
The section of DNA that is desired is cut from the DNA it originated from by
using a restriction enzyme and pasting it into the plasmid by ligation (the
ligase enzyme). Before moving on to transformation, the bacterium is treated
with calcium chloride which allows water to enter the cell and cause it to
swell. When the bacteria swells, it is known as competent bacteria.
The next part of the process is known as transformation. The plasmid with
newly implanted DNA is ready to be inserted into the bacterium. The plasmid
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