BASIC DNA TECHNIQUES AND
GENETIC ENGINEERING
TECHNOLOGY.
INTRODUCTION.
The implementation of genetic engineering on a variety of plants, animals,
and microbes has uses in healthcare, research, industrial, and agricultural
sectors. In scientific study, various organisms undergo genetic
engineering to understand how certain genes operate. In medical or
healthcare sectors , the use of genetic engineering is utilised for treating
infertility, vaccines, various drug therapies and drug discoveries, human
growth hormones, produce insulin, human albumin, antihemophilic
factors, and monoclonal antibodies. Furthermore, converting yeast,
bacteria, insects mammalian cells, or other microorganisms with a gene
encoding a valuable protein has industrial uses. It is possible to produce
huge amounts of the protein by fermenting the transformed organism in
bioreactors and then purifying it. The development of genetically modified
crops and other modified organisms uses genetic engineering as well. The
direct biotechnological change of an organism's genome is known as
genetic engineering, often referred to as genetic modification.
New DNA can be inserted into the host genome once the required genetic
material has been extracted and duplicated using molecular cloning
methods to create a DNA sequence, or by creating a DNA construct,
putting it into the host organism, and then regenerating the DNA. With a
nuclease, genes may be taken out or deleted.
Biology LibreTexts. (2017). 7.23B: Applications of Genetic Engineering. [online]
Available at:
https://bio.libretexts.org/Bookshelves/Microbiology/Microbiology_(Boundless)/
07%3A_Microbial_Genetics/7.23%3A_Genetic_Engineering_Products/7.23B
%3A__Applications_of_Genetic_Engineering#:~:text=In%20medicine%2C%20genetic
%20engineering%20has [Accessed 8 May 2023].
PRACTICAL
1. DNA EXTRACTION FROM BIOLOGICAL SAMPLES.
,DNA carries genetic information inherited from the male and female
parents; this is what is passed on from one generation to another. Except
in RBC due to the absence of the nucleus, the human DNA can be found in
all the cells of the body. DNA extraction can be done from animal and
plant nucleus using certain procedures, chemicals and apparatus. To have
the cell’s DNA extracted, the cell would need to be broken down or lysed
by SDS or washing up liquid. The DNA will need to utilise a suitable
method for extraction, PCR amplification, use restriction enzymes and
undergo fragmentation, then undergo gel electrophoresis for separation
and compare it with fragments of DNA called marker or ladder.
Hypothesis: Except RBC, all eukaryotic cells containing nucleus can have
DNA extraction using appropriate techniques.
Apparatus:
- Beakers
- Glass rod
- Boiling tubes with stoppers
- Boiling tube rack
- Weighing scale
- Pasteur pipette
- Cellophane, stopper or clingfilm
- Measuring cylinders
- Centrifuge
- DNA storage tubes
- Table salt
- Distilled water
- Ethanol
- Buffer solution
Risk Assessment: Alcohol may irritate and the glassware can break. Wear
lab coats, gloves and goggles.
Procedure:
- Label two boiling tubes 1 and 2 and two beakers A and B. then we
take 4 grams of sodium chloride and poured it into beaker A. The
salt was then mixed thoroughly with 46ml of distilled water using a
glass rod to create an 8-to-100 salt solution.
- After that, we put the alcohol on ice. Following that, 1ml of
dishwashing solution was pipetted into boiling tube 1 and secured
with a stopper. We carry out this action because the nuclear and cell
membranes must be lysed, or broken apart, in order to release the
DNA.
, - I next swished water in my mouth approximately ten times to get
some cheek cells. Then, after poring in the water into my beaker B, I
added it to boiling tube 1, which was already filled with the
dishwashing liquid. This is necessary because, if any cheek cells are
present, the washing detergent will go ahead and dissolve the cell
and nuclear membrane, releasing DNA from the cell nucleus.
- The boiling tube 1 containing the cheek cells was then pipetted with
1 ml of our salt solution, and a stopper was placed on it. When water
and sodium chloride are combined, Na+ and Cl- ions are produced.
- Positively charged Na+ ions will bind to any negatively charged
phosphate groups in the DNA to stabilize them and make them
visible when alcohol is added. As a result of the phosphate groups in
its backbone, DNA is negatively charged.
- Then, without shaking the tube since doing so can cause the
washing detergent to generate too much froth or foam, I carefully
mixed everything.
- The solution was then added to boiling tube 1 by carefully pouring
3ml of ice-cold ethanol or alcohol into the cylinder while tilting the
boiling tube. DNA is soluble in water, hence this was done since DNA
cannot be noticed without alcohol. Alcohol does not dissolve DNA,
thus when we put our prepared DNA in alcohol, it condensed and
became visible.
- The boiling tube was then covered for five minutes while we looked
to see whether there was any white substance, which is our DNA
from the cheek cells floating in the tube. To wash the DNA, we then
added 5 ml of ethanol to boiling tube 2.
- Using a Pasteur pipette or glass rod, the DNA was carefully put in
the alcohol that had been prepared in boiling tube 1 to extract it
from the tube 2. Finally, 2ml of distilled water was added to a
storage tube before placing the DNA. Results show the whitish
substance floating and this was the DNA, in the boiling tube.
- The results in conclusion do prove the hypothesis right, as the DNA
was seen as the white strands floating, and this can be seen in the
images above.
Evaluation:
In evaluation, when they used a soft toothbrush, they released about 29
micrograms of DNA as opposed to 17 micrograms when they used a
simple water mouthwash method. Additionally, we also had the option of
GENETIC ENGINEERING
TECHNOLOGY.
INTRODUCTION.
The implementation of genetic engineering on a variety of plants, animals,
and microbes has uses in healthcare, research, industrial, and agricultural
sectors. In scientific study, various organisms undergo genetic
engineering to understand how certain genes operate. In medical or
healthcare sectors , the use of genetic engineering is utilised for treating
infertility, vaccines, various drug therapies and drug discoveries, human
growth hormones, produce insulin, human albumin, antihemophilic
factors, and monoclonal antibodies. Furthermore, converting yeast,
bacteria, insects mammalian cells, or other microorganisms with a gene
encoding a valuable protein has industrial uses. It is possible to produce
huge amounts of the protein by fermenting the transformed organism in
bioreactors and then purifying it. The development of genetically modified
crops and other modified organisms uses genetic engineering as well. The
direct biotechnological change of an organism's genome is known as
genetic engineering, often referred to as genetic modification.
New DNA can be inserted into the host genome once the required genetic
material has been extracted and duplicated using molecular cloning
methods to create a DNA sequence, or by creating a DNA construct,
putting it into the host organism, and then regenerating the DNA. With a
nuclease, genes may be taken out or deleted.
Biology LibreTexts. (2017). 7.23B: Applications of Genetic Engineering. [online]
Available at:
https://bio.libretexts.org/Bookshelves/Microbiology/Microbiology_(Boundless)/
07%3A_Microbial_Genetics/7.23%3A_Genetic_Engineering_Products/7.23B
%3A__Applications_of_Genetic_Engineering#:~:text=In%20medicine%2C%20genetic
%20engineering%20has [Accessed 8 May 2023].
PRACTICAL
1. DNA EXTRACTION FROM BIOLOGICAL SAMPLES.
,DNA carries genetic information inherited from the male and female
parents; this is what is passed on from one generation to another. Except
in RBC due to the absence of the nucleus, the human DNA can be found in
all the cells of the body. DNA extraction can be done from animal and
plant nucleus using certain procedures, chemicals and apparatus. To have
the cell’s DNA extracted, the cell would need to be broken down or lysed
by SDS or washing up liquid. The DNA will need to utilise a suitable
method for extraction, PCR amplification, use restriction enzymes and
undergo fragmentation, then undergo gel electrophoresis for separation
and compare it with fragments of DNA called marker or ladder.
Hypothesis: Except RBC, all eukaryotic cells containing nucleus can have
DNA extraction using appropriate techniques.
Apparatus:
- Beakers
- Glass rod
- Boiling tubes with stoppers
- Boiling tube rack
- Weighing scale
- Pasteur pipette
- Cellophane, stopper or clingfilm
- Measuring cylinders
- Centrifuge
- DNA storage tubes
- Table salt
- Distilled water
- Ethanol
- Buffer solution
Risk Assessment: Alcohol may irritate and the glassware can break. Wear
lab coats, gloves and goggles.
Procedure:
- Label two boiling tubes 1 and 2 and two beakers A and B. then we
take 4 grams of sodium chloride and poured it into beaker A. The
salt was then mixed thoroughly with 46ml of distilled water using a
glass rod to create an 8-to-100 salt solution.
- After that, we put the alcohol on ice. Following that, 1ml of
dishwashing solution was pipetted into boiling tube 1 and secured
with a stopper. We carry out this action because the nuclear and cell
membranes must be lysed, or broken apart, in order to release the
DNA.
, - I next swished water in my mouth approximately ten times to get
some cheek cells. Then, after poring in the water into my beaker B, I
added it to boiling tube 1, which was already filled with the
dishwashing liquid. This is necessary because, if any cheek cells are
present, the washing detergent will go ahead and dissolve the cell
and nuclear membrane, releasing DNA from the cell nucleus.
- The boiling tube 1 containing the cheek cells was then pipetted with
1 ml of our salt solution, and a stopper was placed on it. When water
and sodium chloride are combined, Na+ and Cl- ions are produced.
- Positively charged Na+ ions will bind to any negatively charged
phosphate groups in the DNA to stabilize them and make them
visible when alcohol is added. As a result of the phosphate groups in
its backbone, DNA is negatively charged.
- Then, without shaking the tube since doing so can cause the
washing detergent to generate too much froth or foam, I carefully
mixed everything.
- The solution was then added to boiling tube 1 by carefully pouring
3ml of ice-cold ethanol or alcohol into the cylinder while tilting the
boiling tube. DNA is soluble in water, hence this was done since DNA
cannot be noticed without alcohol. Alcohol does not dissolve DNA,
thus when we put our prepared DNA in alcohol, it condensed and
became visible.
- The boiling tube was then covered for five minutes while we looked
to see whether there was any white substance, which is our DNA
from the cheek cells floating in the tube. To wash the DNA, we then
added 5 ml of ethanol to boiling tube 2.
- Using a Pasteur pipette or glass rod, the DNA was carefully put in
the alcohol that had been prepared in boiling tube 1 to extract it
from the tube 2. Finally, 2ml of distilled water was added to a
storage tube before placing the DNA. Results show the whitish
substance floating and this was the DNA, in the boiling tube.
- The results in conclusion do prove the hypothesis right, as the DNA
was seen as the white strands floating, and this can be seen in the
images above.
Evaluation:
In evaluation, when they used a soft toothbrush, they released about 29
micrograms of DNA as opposed to 17 micrograms when they used a
simple water mouthwash method. Additionally, we also had the option of
