11D
Practical carried out-DNA extraction
Equipment used
- Peas
- Detergent-washing up liquid
- Water
- Mortar and pestle for crushing the peas
- Beaker for preparing the detergent solution
- Stainer
- Test tubes to collect the DNA
- Ice bath to help with the precipitation
- Stirring rod
- Isopropyl alcohol
Method
- Prepare the peas-take a small handful amount of green peas and place them into a clean
mortar and pestle, add a small amount of water to help mash the peas, grind the peas
thoroughly break them down and release the cellular material
- Prepare the detergent solution-in a plastic container add about 50 ml of water with a
teaspoon of the detergent and a pinch of salt(optional)
- Mix this weel until the detergent is dissolved using a glass rod, as the detergent helps
break down the cell membrane and the salt helps the DNA to precipitate.
- Extract the DNA – add the mashed pea mixture into the solution with the detergent
solution and gently swirl it, allow this mixture to sit for few minutes to break open the
plant cells so they can release the DNA into the solution
, - Filter the mixture: Large particles of debris, like solid pea remnants, can be filtered out
using a strainer. Fill a test tube or other sanitized container with the filtered liquid. This
liquid is where the DNA is found. The DNA should precipitate then Fill the test tube with
the filtered liquid, then carefully add the chilled isopropyl alcohol. Slowly add the
alcohol until it separates from the pea solution and forms a layer on top (avoid making it
foamy). The alcohol facilitates the DNA's precipitation from the mixture. the DNA:
White, stringy material should be visible floating at the liquid-alcohol interface. This is a
genetic material. The DNA can be gently collected using a wooden stick or stirring rod.
Bathing in cold water.
PCR experiment- carried out at the university of Salford
Method - Our aim was to amplify a particular DNA segment in the PCR (Polymerase Chain
Reaction) experiment. A DNA sample was first prepared and combined with nucleotides (the
building blocks of DNA), primers (short DNA sequences that flank the target region), a buffer
solution, and DNA polymerase (an enzyme that creates new DNA strands). The reaction mixture
was then put in a thermal cycler, which went through three temperature cycles: extension
(where the polymerase extends the primers to replicate the DNA), annealing (where the
primers bind to the target DNA), and denaturation (where the DNA strands separate). The DNA
was amplified exponentially by repeating this process several times. We acquired a significant
quantity of the target DNA following the cycles, which could then be used to analyze further
experiments.
Gel electrophoresis experiment- carried out at the university of Salford
The DNA fragments we created in the PCR experiment were separated and visualized using the
gel electrophoresis technique. Initially, we made an agarose gel by heating agarose powder and
buffer solution until they melted. After the gel had cooled and solidified in the tray, we added
our DNA samples to tiny wells we had created in the gel. Additionally, to compare the sizes of
our DNA fragments later, we added a DNA ladder—a mixture of known-sized DNA fragments—
to one well. The gel was then exposed to an electric current. Due to its negative charge, DNA
gravitated toward the gel's positive end. Larger DNA fragments passed through the gel more
slowly than smaller ones. After the gel was run, we stained it with a DNA-binding dye and
exposed it to UV light to view the DNA bands. The gel displayed distinct bands in the results.
The size of the amplified DNA fragments could be estimated by comparing the bands from our
DNA samples to the DNA ladder. The bands showed us that we successfully completed the
target DNA, and that the PCR was effective.
,Why are the various steps taken –medical and industrial uses for it.
Making the agarose gel- By serving as a medium, the agarose gel aids in the size-based
separation of the DNA fragments. The agarose forms a gel with microscopic pores when it
dissolves in a buffer solution. Because of this, when an electric current is applied, the DNA
can pass through it.
, Loading the DNA samples-DNA ladder:
The PCR products and a DNA ladder are loaded into the gel's wells. To compare the sizes
of our samples later, the ladder contains DNA fragments of known sizes. By comparing the
amplified DNA fragment's movement through the gel to the ladder's, we can ascertain how
long it is.
Applying the electric current to it: The phosphate groups in DNA's backbone give it a
negative charge. The negatively charged DNA molecules migrate toward the gel's positive
end when an electric current is applied. Because they can more easily pass through the
pores of the gel, smaller DNA fragments move more quickly, causing separation based on
its size.
Staining and visualizing the DNA- Since DNA is colorless, a dye that binds to it and
fluoresces when exposed to UV light is used. As a result, the DNA bands are visible. To
ascertain the existence and dimensions of the DNA fragments, the locations of these
bands can subsequently be examined.
uses Medical uses Industrial uses
1 Genetic testing-the gel Biotechnology and
electrophoresis is used to pharmaceuticals- when
identify specific genetic making biotechnology
markers for diseases it can products, gel
help with mutations in electrophoresis is
genes which cause genetic essential. It is employed in
disorders such as cystic the analysis and
fibrosis. purification of proteins,
enzymes, and other
biological molecules for
use in pharmaceutical
development and
therapeutic interventions.
2 Forensic analysis-is used in Quality control in food and
DNA profiling for criminal agriculture- By examining
investigations e.g. paternity proteins and other
test s and identifying the molecules in the product,
missing persons by gel electrophoresis is used
comparing the DNA in food testing to confirm
samples with each other the existence of genetically
modified organisms (GMOs)
or to guarantee the safety
and quality of food
products.
Practical carried out-DNA extraction
Equipment used
- Peas
- Detergent-washing up liquid
- Water
- Mortar and pestle for crushing the peas
- Beaker for preparing the detergent solution
- Stainer
- Test tubes to collect the DNA
- Ice bath to help with the precipitation
- Stirring rod
- Isopropyl alcohol
Method
- Prepare the peas-take a small handful amount of green peas and place them into a clean
mortar and pestle, add a small amount of water to help mash the peas, grind the peas
thoroughly break them down and release the cellular material
- Prepare the detergent solution-in a plastic container add about 50 ml of water with a
teaspoon of the detergent and a pinch of salt(optional)
- Mix this weel until the detergent is dissolved using a glass rod, as the detergent helps
break down the cell membrane and the salt helps the DNA to precipitate.
- Extract the DNA – add the mashed pea mixture into the solution with the detergent
solution and gently swirl it, allow this mixture to sit for few minutes to break open the
plant cells so they can release the DNA into the solution
, - Filter the mixture: Large particles of debris, like solid pea remnants, can be filtered out
using a strainer. Fill a test tube or other sanitized container with the filtered liquid. This
liquid is where the DNA is found. The DNA should precipitate then Fill the test tube with
the filtered liquid, then carefully add the chilled isopropyl alcohol. Slowly add the
alcohol until it separates from the pea solution and forms a layer on top (avoid making it
foamy). The alcohol facilitates the DNA's precipitation from the mixture. the DNA:
White, stringy material should be visible floating at the liquid-alcohol interface. This is a
genetic material. The DNA can be gently collected using a wooden stick or stirring rod.
Bathing in cold water.
PCR experiment- carried out at the university of Salford
Method - Our aim was to amplify a particular DNA segment in the PCR (Polymerase Chain
Reaction) experiment. A DNA sample was first prepared and combined with nucleotides (the
building blocks of DNA), primers (short DNA sequences that flank the target region), a buffer
solution, and DNA polymerase (an enzyme that creates new DNA strands). The reaction mixture
was then put in a thermal cycler, which went through three temperature cycles: extension
(where the polymerase extends the primers to replicate the DNA), annealing (where the
primers bind to the target DNA), and denaturation (where the DNA strands separate). The DNA
was amplified exponentially by repeating this process several times. We acquired a significant
quantity of the target DNA following the cycles, which could then be used to analyze further
experiments.
Gel electrophoresis experiment- carried out at the university of Salford
The DNA fragments we created in the PCR experiment were separated and visualized using the
gel electrophoresis technique. Initially, we made an agarose gel by heating agarose powder and
buffer solution until they melted. After the gel had cooled and solidified in the tray, we added
our DNA samples to tiny wells we had created in the gel. Additionally, to compare the sizes of
our DNA fragments later, we added a DNA ladder—a mixture of known-sized DNA fragments—
to one well. The gel was then exposed to an electric current. Due to its negative charge, DNA
gravitated toward the gel's positive end. Larger DNA fragments passed through the gel more
slowly than smaller ones. After the gel was run, we stained it with a DNA-binding dye and
exposed it to UV light to view the DNA bands. The gel displayed distinct bands in the results.
The size of the amplified DNA fragments could be estimated by comparing the bands from our
DNA samples to the DNA ladder. The bands showed us that we successfully completed the
target DNA, and that the PCR was effective.
,Why are the various steps taken –medical and industrial uses for it.
Making the agarose gel- By serving as a medium, the agarose gel aids in the size-based
separation of the DNA fragments. The agarose forms a gel with microscopic pores when it
dissolves in a buffer solution. Because of this, when an electric current is applied, the DNA
can pass through it.
, Loading the DNA samples-DNA ladder:
The PCR products and a DNA ladder are loaded into the gel's wells. To compare the sizes
of our samples later, the ladder contains DNA fragments of known sizes. By comparing the
amplified DNA fragment's movement through the gel to the ladder's, we can ascertain how
long it is.
Applying the electric current to it: The phosphate groups in DNA's backbone give it a
negative charge. The negatively charged DNA molecules migrate toward the gel's positive
end when an electric current is applied. Because they can more easily pass through the
pores of the gel, smaller DNA fragments move more quickly, causing separation based on
its size.
Staining and visualizing the DNA- Since DNA is colorless, a dye that binds to it and
fluoresces when exposed to UV light is used. As a result, the DNA bands are visible. To
ascertain the existence and dimensions of the DNA fragments, the locations of these
bands can subsequently be examined.
uses Medical uses Industrial uses
1 Genetic testing-the gel Biotechnology and
electrophoresis is used to pharmaceuticals- when
identify specific genetic making biotechnology
markers for diseases it can products, gel
help with mutations in electrophoresis is
genes which cause genetic essential. It is employed in
disorders such as cystic the analysis and
fibrosis. purification of proteins,
enzymes, and other
biological molecules for
use in pharmaceutical
development and
therapeutic interventions.
2 Forensic analysis-is used in Quality control in food and
DNA profiling for criminal agriculture- By examining
investigations e.g. paternity proteins and other
test s and identifying the molecules in the product,
missing persons by gel electrophoresis is used
comparing the DNA in food testing to confirm
samples with each other the existence of genetically
modified organisms (GMOs)
or to guarantee the safety
and quality of food
products.
