Extraction Of DNA In Tomatoes And Garden Eggs Fruits

Project and Seminar Material for Science Laboratory Technology SLT

Extraction Of DNA In Tomatoes And Garden Eggs Fruits


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
  • Certification
  • Dedication
  • Acknowledgement
  • Abstract
  • Table of content

Chapter One

1.0 Introduction

  • 1.1 History of DNA
  • 1.2 Definition of DNA
  • 1.3 Definition of extraction of DNA
  • 1.4 Importance of DNA extraction
  • 1.5 The human Genome project
  • 1.6 How genes are sequence

Chapter Two

2.0 Literature Review

  • 2.1 Concept of DNA replication
  • 2.2 Eukaryotic DNA replication
  • 2.3 Protein associated with DNA replication and phase keeping

Chapter Three

3.0 Materials and Methodology

  • 3.1 Collection of the samples
  • 3.2 Preparation of the samples
  • 3.3 Procedure
  • 3.3.1 Preliminary operations
  • 3.3.2 Preparation of the extraction solution
  • 3.3.3 Preparation of the Pulp
  • 3.3.4 Filtration of the Pulp
  • 3.3.5 Extracting of DNA
  • 3.3.6 Extracting of DNA

Chapter Four

4.0 Result


Chapter Five

  • 5.0 Discussion
  • 5.1 Conclusion
  • 5.2 Recommendation
  • References

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).


Chapter Five


Discussion

The DNA is of immense importance that each organism has its peculiar DNA and it has been conformed over time that no two organisms possess the same DNA.

This has a great benefit in identification and classification of organism, both plants and animals and even in micro-organism. The study of DNA is a current one in the field of biological sciences.

Therefore, the method used in the extraction of DNA and their isolation is current simple and cheap experiment which can be prepared both at home and the school laboratory.

The helical structure observed on the shape of isolated DNA of these two fruits confirmed previous research work that DNA is a double stranded organic molecule and that is helical in shape. (Sylvia J, 2001).

The analysis of the isolated DNA was supposed digestion of DNA molecule using restriction enzymes which is responsible for cutting DNA into different specific segments so that different sized fragments will result.

The next step after this is supposed to be the remaining of the segmented gel fragment on the gel electrophoresis so that it will function in separating the different DNA fragments according to their length. This is because shorter fragment migrate further in an electric field than do longer fragments.

The fragments are denatured (separated) and transferred to a membrane where a radioactive probe can apply.
Lastly, the resulting pattern (separate) and transfer to a membrane where a radioactive probe can be detected by an autoradiography. (Boyer B, 1986).

Unfortunately, the aspect of DNA analysis could not be carried out due to lack of facilities like the gel electrophoresis, a radioactive probe and auto-radiography and the DNA molecule restriction enzyme. Efforts made by carrying out analysis could not be done due to the time and financial constraint. The problem majorly is the absence of the enzymes in the two centers. Hence, the enzymes would have been import from the United Kingdom and aside from this, a very conducive environment with regulated temperature under regular power supply is very essential, this is because these enzymes thermally labile. The polytechnic can take a held step by providing all the necessary equipments and enzymes so that it could serve as a main centre in Nigeria of income generated for the polytechnic. (Boyer J, 1986).


Conclusion

On the basis of he results obtained in this study, the following conclusion can be made that the extraction of DNA in tomatoes and garden egg is cheep and easy to carry out using this method for any types of fruits or vegetables which can be use to detect bacterial and viruses in the environment as well as diagnosing disease and genetic disorders.


Recommendation

This project is highly recommended to students by try to practicalize it which helps in identification and classification of species of plant families also to determine how they are related.


Extraction Of DNA In Tomatoes And Garden Eggs Fruits


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Extraction Of DNA In Tomatoes And Garden Eggs Fruits


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