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Mutagenesis Lab

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The purpose of this lab was to observe bacterial mutagenesis of E. Coli by ultraviolet (UV) induced mutation, and observe these mutations with the use of DNA isolation techniques and gel electrophoresis vs. a control. A mutation is the changing in sequence of nucleic acids (1). When restriction endonucleases are added to DNA, the enzyme will read the certain sequence of base pairs that correlate to that enzyme of use, and then will cleave the DNA at the restriction site (2). In mutagenized DNA, the enzyme will be unable to recognize the sequence in which to cut the DNA, and this could be observed with the use of gel electrophoresis by observing the banding patterns of a control vs. the mutagenized (2). The mutagenized may also have less total…show more content…
When choosing the restriction enzymes to use, many factors are considered such as methylation of certain genes, which can influence the results of the experiment. Methylation will protect the gene from restriction enzymes, so in this experiment restriction enzymes that are not methylation sensitive are used (3). The wavelength of the UV Radiation can also have an impact on the rate of mutagenesis (4), which will be discussed more in the results. Restriction enzymes in this experiment such as Bam will cleave every six base pairs at a specific location on the target site (5). Bacteria such as E. Coli can naturally make restriction enzymes such as Eco, which is used naturally by the host cell to cut viral DNA (typically from phages) and make it non-functional in order to protect the integrity of its genome (6). In this discovery-based experiment, we will observe the changes in banding patterns of E. Coli DNA with multiple restriction endonucleases after UV induced mutation, and use these results against the control to determine if there are any changes in gene sequence by the number of base pairs in the observed DNA
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