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Extraction of plasmid DNA and set up of polymerase chain Reaction (PCR) - Coursework Example

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It basically amplifies a particular gene by conducting an in vitro DNA replication. For this process to proceed, the necessary ingredients, such as the appropriate…
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Extraction of plasmid DNA and set up of polymerase chain Reaction (PCR)
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"Extraction of plasmid DNA and set up of polymerase chain Reaction (PCR)"

Download file to see previous pages g temperatures to different levels, the different steps of DNA replication, the separation of the double strand, the annealing of primers to the DNA single strands, and the activity of DNA polymerase are facilitated. The cycle of changing temperatures are repeated, producing many strands in the process. Of course, the DNA polymerase should be able to withstand these changes in temperature.
The resulting amplified gene can subsequently be characterized, either through identification, measurement or expression. The information obtained from this can further be used to characterize the resulting protein product or the organism from which it was obtained. Due to the vast amount of processes PCR products can undergo, the technique has been the staple for laboratory diagnosis of diseases, bacteria and virus identification, crime scene investigations, and others. As can be seen, PCR follows and precedes many other molecular processes.
In this experiment, plasmids from a transformed bacterial cell pellet were extracted to isolate the gene encoding for red fluorescent protein (RFP). After measuring the amount of DNA extracted, it was then prepared for PCR.
Bacterial transformation, or insertion of gene of interest inside bacteria cells, is a viable strategy in not only housing the gene, but also amplifying the sequence. Thus, bacterial strains used for this purpose, such as E. coli DH5α, JM109 and XL-1 Blue, are specialized not only to be able to imbibe the plasmids upon electric or chemical induction, but also to replicate the inserted genetic material during cell division (Yoshida and Sato, 2009).
On the other hand, special plasmids, called vectors, are circular DNA used to accompany the gene inside the bacterial cell. It contains sequences for restriction enzymes, so that the vector circular configuration can be broken into two parts, through which the gene of interest can be inserted into, re-establishing the circular appearance of the vector. The vector also ...Download file to see next pagesRead More
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