ABSTRACT
This
project is basically concerned with the extraction of DNA in tomatoes and
garden egg fruit.
An
experiment on how to extract DNA on tomatoes and garden egg, isolation on it to
see what DNA actually look like as well as importance of DNA extraction are
consid3ered in this project.
TABLE
OF CONTENT
Title page ……………………………………………………… i.
Certification……………………………………………………..ii
Dedication……………………………………………………….iii
Acknowledgement……………………………………………..iv
Abstract………………………………………………………….v
Table of content……………………………………………….vi
CHAPTER
ONE
1.0
Introduction………………………………………………1
1.1
History
of DNA…………………………………………..1-3
1.2
Definition
of DNA……………………………………….6
1.3
Definition
of extraction of DNA……………………….6
1.4
Importance
of DNA extraction……………………7
1.5
The
human Genome project………………………7-12
1.6
How
genes are sequence……………………………..12
CHAPTER
TWO
2.0
Literature
review………………………………………13
2.1
Concept
of DNA replication…………………………13
2.2
Eukaryotic
DNA replication………………………13-17
2.3
Protein
associated with DNA replication and phase keeping…………………………………………………18-23
CHAPTER
THREE
3.0
Materials
and methodology……………………………24
3.1
Collection
of the samples……………………………..25
3.2
Preparation
of the samples………………………25-26
3.3
Procedure………………………………………………26
3.3.1 Preliminary
operations…………………………………27
3.3.2 Preparation
of the extraction solution……………..27
3.3.3 Preparation
of the Pulp………………………………..28
3.3.4 Filtration
of the Pulp…………………………………...28
3.3.5 Extracting
of DNA……………………………………….29
3.3.6 Extracting
of DNA………………………………….29-34
CHAPTER
FOUR
4.0
Result…………………………………………………35-36
CHAPTER
FIVE
5.0
Discussion……………………………………………37-39
5.1
Conclusion……………………………………………..40
5.2
Recommendation………………………………………41
References……………………………………………42-44
CHAPTER
ONE
1.0 INTRODUCTION
1.1 HISTORY OF DNA
THE
STRUCTURE OF DNA
Working
together at the University of Cambridge in England, James Watson, An American
Scientist and Francis Crick, a British researcher made a major scientific
breakthrough, when they discovered the famous “double helix”. The structure of
DNA, the molecule of life.
In
April 25, 1983, issue of science Journal Nature, Watson and Cricks Wrote, we
wish to suggest a structure for the slat of dexyribose Nucleic acid (DNA). This
structure has novel features which are of considerable biological interest.
Nine
years later, in 1962, they received the Nobel prizes for answering one of science’s
long pondered mysteries, advancing the emergency field of molecular biology in
the process.
Watson
and Crick’s quest helps illustrate how collaboration, creativity, handwork and
serendipity often conspire on the oath to scientific achievement.
A EUREKA MOVEMENT
Decyribonecliec
acid (DNA) was first isolated in 189 by the Swiss scientist friedrish Mieschre
he called the white, slightly acidic chemical that he fond in cells “nulein”.
By the late 1940s, scientists knew what DNA contained-phosphate sugar and four
nitrogen containing chemical “bases” adenime (A), thymine (T), Guanine (G), and
cytosine (C). But no one has figured out what the DNA molecule looked like.
Friedrich (1869).
In 1993, Linus Pauling, the great
American Chemist, claimed to have discovered the structure of the DNA molecule,
but when Watson saw Pauling’s research paper (which has not yet been published)
on January 28, 1953, he knew it was wrong. A few days later at King’s college
in London,
Watson was shown an x-ray diffraction photograph of the DNA crystal taken by
scientist (Watson J, 2001).
WHAT
IS DNA?
DNA stands for deoxyribonucleic acid. It is a
long polymer-type molecule in which the replicating groups are nucleotides.
Deoxyribonucleic acid, more commonly known as
DNA is a complex molecule that contains all of the information necessary to
build and maintain an organism. All living things have DNA within their cells. Infact,
nearly every cell in a multi-cellular organism possess the full set of DNA
required for that organism. However, DNA does more that specific the structure
and function of living things. It is also serves as the primary unit of
heredity in organism of all types. In other words, whenever organisms of all
types in reproduce, a portion of their DNA is passed along to their offspring.
This transmission of all or part of an organism’s DNA helps ensure a certain
level of continuity form one generation to the next, while still allowing for insight
changes that contribute to the diversity of life.
Most DNA is located in the cell nucleus where
it is called nuclear DNA but a small amount of DNA can be founded in the
mitochondria where it is called mitochondria DNA (Rasmussim S, 2000).
DNA which is a long molecule, like a chain
where the link of the chain are pieces called nucleotide some times also called
“bases”. There are four different types of nucleotides in DNA which is called
“A” “G”, “C”, and “T”. These four are all that is necessary to write acode that
describes our entire body plan. The four nucleotides look a little bit alike. They
all have a ring of carbons called, in chemist’s terminology, a ‘sugar’.
However, each nucleotide also has another type of ring structure and this is
where the four types of nucleotide are different. These rings are organic base
much like the more familiar mineral acids and bases like NOH or HCL, except
these bases are composed of carbon, nitrogen and oxygen. (Rasmiussim S, 2005).
EXTRACTION
OF DNA
Since
DNA is the blueprint for life, everything living contains DNA. DNA isolation is
one of the most basic and essential techniques in the study of DNA. The
extraction of DNA from cells and its purification are of primary importance to
the field of biotechnology and forensic. Extraction and purification of DNA are
the first steps in the analysis and manipulation of DNA that allow scientist to
detect genetic disorder, produce DNA finger prints for individuals and even
create genetically engineered organisms that can produce beneficial products
such as insulin, antibiotics and hormones. (Kelly O, 2004).
WHAT
IS DNA EXTRACTION?
DNA Extraction is the removal of deoxyribonucleic
acid (DNA) from the cells or viruses in which it normally resides.
WHAT
IS IT USED FOR?
Extraction
of DNA is often an early step in many diagnostic processes used to detect
bacteria and viruses in the environment as well as diagnosing disease and
genetic disorders.
THE
HUMAN GENOME PROJECT
This is an international human genome
sequencing consortium that was forms in (1990) with the goal of indenting 3
billions bits (base pairs) of DNA that comprises the entire human genetic code
in the genome and put them in proper sequences. The consortium consists of some
20 groups from the United State, United Kingdom, France,
Germany, Japan and China,
funded mainly by the .S National Institutes of Health (NIH) and the welcome
trust of London,
to the tune of about 2.5 billions.
The
goal was originally to finish assembling the human genome by 2003. The
consortium laboratories sequence the genome one bit at a time, very
methodically, “Clone-by-done” sequencing (Dr. Craig, 1998).
Importance
of DNA extraction is often an early step in many diagnostic processes used to
detect bacteria and viruses in the environment as well as diagnosing disease
and genetic disorders.
This
techniques involved fluorescence in situ hybridization (fish); fish is a molecular
techniques that is used among other things to identity and enumerate specific
bacteria groups sequencing position of whole genomes may be sequenced as well
as extra chromosome element for comparison with existing sequences in the
public data base.
How
are gene sequenced? Gene sequencing is a complicated, but now fairly routine,
process. Overlapping fragments of genes are sequenced, and from the overlapping
portions, the entire gene can be deduced. The process involves first isolating
and multiplying (cloning) large segments of the genes to provide enough to work
with. These cloned segments constitute a genomic library and are provided to
sequencing laboratories. The long overlapping segments are broken into shorter
splices, using a random shearing technique in which the sample is forced
through a needle and the shear forces break them up. Originally, they were
spliced using a nuclease enzymes, but the shearing techniques gives more randomness.
(Nucleases degrade nucleic acids. They may act on only single strands or on
double stands, or on both. A nuclease may degrade from one end, an exonuclease
or begin in the middle-an endonuclease). The splices are replicated by pairing
with the appropriate complementary nucleotides. Beginning at one end, one nucleotide
at a time is added. Ordinarily, this would gomuti het entire fragment is
replicated. But, we have a way of randomly stopping the replication at
different points along the DNA, for each of the four nucleotides. We also have
a way to measure what the terminating nucleotide is, whether it is A, C, G, or
T. if among the millions or billions of fragment clones, we obtain some of very
possible length along the nucleotide sequence, and if we can separate them by
length and line then up, we will have to complete ordered set or nucleotide in
the original DNA template. (Frederick S, 1970).
The
century of the gene began when James Watson and Francis Crick unravels the secret
of the building block of genes-DNA exist as a double helix. It took the next 40
years to actually determine the entire human gene sequences (the genome).
The
real building blocks of our body are proteins. Nearly all biological activity
is carried out by proteins. Our muscles, testis, liver, skin blood just b about
all of our bodies use proteins to create the chemistry of life. Genes make up
our chromosome. The nucleus of nearly every one of our 100 billions cells
contains 23 pairs of chromosomes that is 46 individual chromosomes. Exceptions are
sperms and an eggs cell, each contains 23 chromosomes. Each parent contributes
23 chromosomes to a child. Each child gets only a portion (a subset) of each
parent’s DNA and the pieces received are random so each child gets a different
subset of each parent genes except for identical twins, which have identical
genes.
Chromosomes, when observed under a
microscope, are fatty, tabular-looking substances. They are infact made up of a
series of genes, which are tightly twisted double strands of DNA molecules. The
two strands are held together by weak hydrogen bonding between pairs of nucleotide
bases on each strained, which a may be viewed as “steps” in the DNA “ladder” (Harrison
J, 2000).
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