SURNAME : MASONDO

NAME : BENSON LETHUKUTHULA

STUDENT NUMBER : 202126087

DATE : 12 MAY 2023

COURSE CODE : BCH 212

EXPIREMENT NUMBER : 7

EXPIREMENT TOPIC : isolation of DNA from onion

Title : isolation of DNA from onion

INTRODUCTION:

Gel electrophoresis is the standard lab procedure for separating DNA by size (e.g., length in
base pairs) for visualization and purification. Electrophoresis uses an electrical field to move
the negatively charged DNA through an agarose gel matrix toward a positive electrode.
Shorter DNA fragments migrate through the gel more quickly than longer ones. Thus, you
can determine the approximate length of a DNA fragment by running it on an agarose gel
alongside a DNA ladder (a collection of DNA fragments of known lengths)(
Timms,Cramer,2008).

DNA, or deoxyribonucleic acid, is the genetic material that carries the instructions for the
development and function of all living organisms. This complex molecule is found in every
cell of the body, and its structure and function have been studied intensely for many years by
scientists across the globe. DNA is composed of four different nitrogen-containing bases
(adenine, guanine, cytosine, and thymine), a sugar molecule, and a phosphate group. It is
arranged into a double helix structure, with the bases pairing up in a specific way (A with T,
and C with G). The sequence of these bases determines the genetic code, which is responsible
for the traits and characteristics of an organism.DNA replication is the process by which cells
create copies of their DNA, in order to pass genetic information on to the next generation.
This process is essential for cell division and growth, and is controlled by a complex network
of enzymes and proteins. However, mutations can occur during replication, which can lead to
changes in the genetic code and the development of genetic diseases(Lacks, Hotchkiss,
1960).

The study of DNA has revolutionized many fields of science, from medicine to forensics to
evolutionary biology. Researchers can use DNA analysis to identify individuals, diagnose
genetic disorders, and track the evolution of species over time. Advances in DNA sequencing
technology have allowed scientists to map the entire human genome, which has opened up
new avenues of research into the causes of disease and potential treatmentsDNA is a
fascinating and complex molecule that plays a central role in the development and function of
all living organisms. Its study has led to many important advances in science, but also raises
important ethical questions about privacy and discrimination. As research in this field
continues to advance, it is important to carefully consider the potential implications and
ethical challenges of DNA analysis(Kabanov,Kabanov,1995).
Deoxyribonucleic acid (DNA) is the genetic material found in all living organisms, which
contains the instructions for the development, function, and reproduction of all living
organisms. Therefore, understanding the structure and function of DNA is essential to
understand the fundamental processes of life. One of the critical steps in studying DNA is
extracting it from cells or tissues. The isolation of DNA from cells is crucial for many genetic
studies, including identifying genetic disorders and developing new vaccines and drugs.There
are different methods for isolating DNA, each with its advantages and disadvantages. The
most commonly used method involves the extraction of DNA using a combination of
mechanical and chemical methods. Mechanical methods involve breaking cells through
physical force, such as grinding in a mortar or using a homogenizer. Chemical methods
involve the use of reagents that solubilize cell membranes and protein molecules that
surround DNA. These methods are usually less harsh and more effective than mechanical
methods(Jie et al, 2003).

The isolation of DNA is typically performed in several stages, depending on the type of tissue
or cell that is being used. The first stage involves the disruption of the cell membrane to
release the contents of the cell, including the DNA. This stage is typically done using a buffer
solution that contains salts, detergents, and enzymes that break down the structure of the cell.
The next stage involves the removal of proteins from the DNA, which can be done using an
enzymatic digestion or a salt-based precipitation. The final stage involves the purification or
concentration of the extracted DNA, which can be done using different techniques such as
ethanol precipitation or chromatography(Hadke, 2003)

There are several factors to consider when isolating DNA, including the type of tissue, the
amount of DNA required, and the downstream applications. For example, different types of
tissues require different methods of DNA extraction, as some are more challenging to break
down than others. Additionally, the amount of DNA required for different applications can
vary, and some methods may be more efficient at recovering larger amounts of DNA than
others. Finally, some downstream applications may require specific DNA qualities, such as
high purity, high molecular weight, or minimal contamination.The isolation of DNA is a
critical step in studying its structure and function. Several methods are available for isolating
DNA, each with its advantages and disadvantages. The choice of method depends on several
factors, including the type of tissue, the amount of DNA required, and the downstream
applications. Advances in technology and understanding of the mechanisms behind DNA
extraction have improved the efficiency and effectiveness of DNA extraction methods,
providing researchers with high-quality DNA samples for genetic studies(Di Pinto,2007).

AIM: The aim of the practical was to isolate DNA from an onion and to discover the
fundamental properties of DNA and scientific methods required to isolate it.

METHOD AND MATERIAL:

1.Isolation of DNA

80 ml of water, 10 ml of dish washing detergent alongside the non-iodized salt were

additional to a 250 ml of a beaker. 50g of finely grated onion was added on a high of the
mixture. The whole answer was incubated in 60 degrees Celsius for 15 minutes. Then the
material was then filtrated through a layer of cheese textile and filtrated answer was
contained with the presence of the deoxyribonucleic acid. 1g of meat tenderizer was added to
a strain of the material and unbroken for 5 min in temperature. A 50 ml ice isopropanol was
added and poured on the facet ways of the tipped beaker, the alcohol and material fashioned
very different layers with material on bottom. White string of deoxyribonucleic acid was
spooled out with a glass rod because the rod was turned on an equivalent direction. The
deoxyribonucleic acid (DNA) was re-suspended into a five small litre TAE buffer

2.Preparation of 0.4 agarose gel

stock solution of 5ml was added to a 495ml of distilled water ,the solution was mixed in a
reagent bottle to make a buffer solution .The solution was then transferred to an Erlenmeyer
flask .The solid agarose powder of 0,4g was then added to the solution in the flask .The
solution is then heated in the microwave oven ,buffering steps were repeated and the buffer
was added to the same reagent bottle on top of the remaining buffer mixture .Pipette of 0,5µl
of ethidium bromide was then used placed inside the agarose gel or solution then waited in
the casting trays

Discussion:

In this practical onion was used which was the good idea because onions are good for DNA
extraction experiments because they contain a high amount of DNA in their nuclei. The DNA
is also less degraded in onion cells compared to other plant tissues. Onion cells are also easy
to break open and release their DNA. In addition, onions do not contain enzymes that degrade
DNA, making it easier to extract the DNA without damaging it. Therefore, onion extract is
commonly used as a source for DNA isolation in laboratory experiments.

Salt is added to the solution when isolating DNA to create an isotonic environment. This is
important because DNA is insoluble in alcohol, which is used to precipitate it out of solution.
By adding salt, the charged sodium ions in the salt compete with the phosphate groups on the
DNA molecule, reducing the amount of repulsion between the phosphate groups. This makes
the DNA molecule more compact and helps it to more easily precipitate out of solution when
alcohol is added. Salt also helps to neutralize the charge of the DNA, making it more stable
and less likely to degrade. Overall, adding salt to the solution when isolating DNA helps to
increase the yield and purity of the isolated DNA. If you cannot find dishwashing liquid, you
can also use other detergents such as sodium dodecyl sulfate (SDS) or Triton X-100.
However, these detergents may require different extraction protocols and their efficiency may
vary depending on the type of sample being used. It is always important to follow the specific
protocol for the DNA extraction method being used.

The function of meat tenderizer is to break down the proteins in meat, making it more tender.
This is achieved by using proteolytic enzymes such as papain, bromelain or actinidin. These
enzymes break down the tough muscle fibers, making the meat more palatable.In DNA
isolation protocols, commercial reagents are often used to break down cell membranes and
nuclear membranes to release DNA from cells. One such reagent is Proteinase K, which is a
broad-spectrum serine protease that cleaves peptide bonds in proteins. Proteinase K is often
preferred over meat tenderizer since it specifically targets proteins without affecting DNA,
and it is highly efficient in lysing cells and releasing DNA.
RESULTS

Figure 1:Agarose gel electrophoresis image with 10 wells(stained with ethidium bromide)
REFFERENCES

Di Pinto, A., Forte, V., Guastadisegni, M.C., Martino, C., Schena, F.P. and Tantillo, G., 2007.
A comparison of DNA extraction methods for food analysis. Food control, 18(1), pp.76-80.

Hadke, S.P., Wayal, S.R. and Londhe, N.B.,2003, ISOLATION OF DNA FROM ONION.

Jie, L., Chengri, C. and Jingpeng, L., 2003. A Simple and Rapid Method for the Isolation of
Mitochondrial DNA from Onion. Acta Horticulturae Sinica, 30(5), p.553.

Kabanov, A.V. and Kabanov, V.A., 1995. DNA complexes with polycations for the delivery
of genetic material into cells. Bioconjugate chemistry, 6(1), pp.7-20.

Lacks, S. and Hotchkiss, R.D., 1960. A study of the genetic material determining an enzyme
activity in pneumococcus. Biochimica et biophysica acta, 39(3), pp.508-518.

Timms, J.F. and Cramer, R., 2008. Difference gel electrophoresis. Proteomics, 8(23‐24),
pp.4886-4897.