The Community of Microorganisms that Reside in the Epithelia of Humans

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There is a large number of species of microbes found on the human body. This bacterial organism are found in the skin, mouth, or nose. This lab consisted of the collection of skin bacterial organisms and amplification of the 16s rRNA to construct a small molecular phylogeny of the human body microbiome, or the community of microorganisms that reside in the epithelia of humans. This information could only be acquired through processes such as DNA extraction, amplification of specific genetic target by the polymerase chain reaction (PCR), agarose gel electrophoresis, restriction enzyme digestion, cloning of DNA fragments into plasmid vector, transformation and blue/white clone colony screening. Through the phylogenetic tree analysis…show more content…
coli are picked and the plasmids are purified. The purified plasmids are used as a template for the sequencing reaction. The objective of the lab was to learn how to use the polymerase chain reaction (PCR) to amplify the small subunit ribosomal RNA gene from a bacteria colony, also be able to run an agarose gel to visualize the resulting PCR amplifications and extract the amplified DNA from the agarose gel. Methods/Materials There were several steps used to acquire the colony necessary for the PCR. First a student forearm was swabbed using a cotton swab, the cells were then placed in an agar plate. DNA was then extracted from the cultured bacteria by using a technique to lyse the cells and solubilize the DNA, then enzymes were used to remove contaminating proteins. The DNA extraction consisted of a lysis buffer that contained high concentrations of salt for denaturing. Binding with the use of ethanol and a washing step to purify the DNA. The final step for the DNA extraction was elution where the pure DNA was release. Proceeding the extraction of DNA the results of the 16s gene amplification were examined through gel electrophoresis it was analyzed by estimating the size of the PCR bands with marker bands. After measuring the success of the extraction, a technique called TA cloning was started. Cloning of PCR products was done by using partially purified amplified products with

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