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Recombinant Dna Lab Report

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Preparation of Recombinant Intermediates; Topologically different forms of DNA

INTRODUCTION
The gene is the cornerstone of the Molecular Biology techniques. These genes can be isolated and amplified for the better study. One of the most important methods in Molecular Biology is the insertion of desired gene or gene of interest into a vehicle or vector that can be replicated in living cells. This process is called cloning. The result of these two DNAs combination is called recombinant DNA molecule. Crossing-over which is a genetic process will produce recombinant DNA normally. This term is given for the DNA molecule produced by joining the segments derived from different biological sources. Host cell is the one which is the place to keep these recombinant DNA molecule and is prokaryotic or eukaryotic (mostly prokaryotic). When the host cell replicates, the piece of DNA within the vector will be get replicated. Thus, got the amplified foreign DNA, which is numerous in number and can be purified and for the further analysis (Kary B Mullis, 1990)
Geneticists use recombination term to describe the outcome of crossing over …show more content…

There are three different bacterial systems present. RecE, RecF, and RecBCD are the bacterial systems. The RecBCD is most important in bacteria. RecBCD enzyme will initiate the recombination. Enzyme is also called as exonuclease V, which have both nuclease and helicase activity. The enzyme complex have three subunits; RecB, RecC, and RecD. It is a bipolar DNA helicase enzyme. RecB and RecD have ATP-dependant helicase activity. RecB travel on 3'→5' direction on one strand and RecD on the other strand on 5'→3' direction. RecC recognizes specific sequence in DNA, called chi (Crossover Hotspot Instigator) site (Pranav Kumar and Usha Mina,

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2. How does DNA replicate itself? In order to replicate itself, DNA undergoes DNA replication, a process in which the DNA unwinds and splits in two. From that point on, new nucleotides are added to each of the original strands (A to T, C to G) until the result is two identical sequence copies of DNA.

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The same region is also amplified on both chromosomes, however they are different sizes, which are then put into gel

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Courtney Peters Chapter 6: Finding mutant phenotypes for cloned genes 1. “Reverse genetics” refers to the process of finding a mutant phenotype for a gene whose DNA sequence is known. a. What are some of the situations in which the DNA sequence of a gene, and perhaps evidence about its function, can be known before the mutant phenotype is known? Gene expression can be used to find out the function of a gene before its mutant phenotype as well as genetic screening. b.

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The plasmid and the gene are both cut using restriction enzymes. To incorporate the gene into the plasmid, both the plasmid and gen stuck together by a bacterial enzyme called ligase. The plasmid with the foreign gene can then be inserted into the bacteria One specific plasmid is pUB110 which is circular whose host is Bacillus subtilis. It has a plasmid size 2.3 kpb and copy number between 20 - 50. We can selectively grow bacteria that contain this plasmid by using selective and differential media.

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This experiment undergoes the use of recombinant DNA technology. This is the merging of DNA molecules. The molecules are extracted from two diverse species which are then introduced into a host organism. This then creates a first-hand genetic arrangement which can be of worth to science, the medical field, cultivation and diligence. It is generally straight forward to segregate an example of DNA from an assortment of cells (Telser, 2002).

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These expressions of thought are ambiguous to the reader, which is disappointing since the scientific explanations of genetic transfer were explained in clearly. Although lacking creative writing style, the article provides effective visual aid for a teen audience to be engaged and inquiring to learn more about the issue. The diagram of a bacterial cell offers readers a comparison of bacterial chromosomes with that of plasmids. The cell does not include any other organelles to confuse or distract the student.

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The single crossovers have frequencies that are in between the parental and the double cross overs. Recombinant frequencies are important for determining whether genes are linked or not. In Prophase I of meiosis, homologous chromosomes exchange information in a process that is called crossing over. This process creates the recombinant genes that

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Genetic Engineering, variously know as genetic modification or genetic manipulation, is the set of tools used to manipulate the genes of an organism. Genetic Engineering (GE) is different from the traditional breeding, where the organism’s genes are modified indirectly. In fact, G.E. uses the technique of molecular cloning and transformation to alter the structure and characteristics of genes directly. Genetic Engineering comes under the broad heading of biotechnology and it still remains a highly controversial debate based on the fact that it raises moral dilemmas. Basically, Genetic Engineering refers to the reshuffling of genes, usually from one species to another.

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Changes introduced in an animal’s genetic structure can therefore be transmitted to the next generation. How is this done? Gene transfer technology is an advanced version of a cut-and-paste operation. Once the wanted gene is identified in the organism's genome, it can be taken out, transferred to a plant, and pasted into its genome. Once the new gene has been introduced, the plant can be bred to create a new strain that passes the gene from generation to generation (The Genetic Science Learning Center at the University of Utah,

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Viruses are the ideal mechanism for gene transfers because of their capability of injecting their own genetic material into a patient's genome as a means of reproduction (Lyford J.). Before doctors can begin the procedure, they must be able to identify the specific gene on a chromosome that is responsible for causing the disorder; they verify this through genetic testing. In genetic testing, a strand of DNA with restriction enzymes is cut and inserted into plasmid. It is then analyzed to locate the gene. After this process is completed scientists are able to begin strategizing their plans for gene therapy (Bergeson.).

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The altering of human DNA with genetic engineering, as well as the replication of human

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In this process, a nucleus is removed from an egg. Next, a cell from the individual who is being cloned is placed into

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The process of creating a transgene begins by selecting the gene of interest from an organism or selecting for purchase any of the thousands of known genes from massive online genomic databases. The field of transgenics allows scientists to develop organisms that express a trait not normally found in a species. An example of this would be hypoallergenic pets that people with bad allergies can have as pets without all the negative effects of the allergens that come with a regular animals. Nowadays, breakthroughs in molecular biology are happening at an unprecedented rate. One of them is the ability to engineer transgenic animals for example animals that carry genes from other species.

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The definition is simple, you are essentially removing the bad part of the gene and replacing it with a healthier, new model. We do this to try and correct the genetic parts that are causing the problems within the cells, thus preventing the disease all together.(3) Now, you cannot just go and split the cell in half and poke around, you have to enter the cell using what is called a vector. Most common vectors used in today’s research are viruses.

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Genetic Engineering is a process where we manually remove our gene of interest from an organism and insert it into another organism. This results in the organism possessing the particular trait we want from the organism. Applications of genetic engineering can be found all around us. They can range from certain crops that are pest resistant to the food with higher nutritional value and even to the medicine that we consume. Genetic engineering began in 1973 when Herb Boyer and Stanley Cohen created the first recombinant deoxyribonucleic (DNA) organism.

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