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Recombinant DNA Method- Mutagenesis By: Jessica Green
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Mutagenesis Mutagenesis (the creation or formation of a mutation) can be used as a powerful genetic tool. By inducing mutations in specific ways and then observing the phenotype of the organism the function of genes and even individual nucleotides can be determined.
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Mutagenesis Laboratory
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Some types of Mutagenesis Directed Mutagenesis Site-directed/Site-specific Mutagenesis Mismatched Mutagenesis
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Directed Mutagenesis a largely discredited hypothesis proposing that organisms can respond to environmental stresses through directing mutations to certain genes or areas of the genome.
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Site-directed Mutagenesis Where a specific site in a cloned DNA needs to be altered in a precise, pre- determined way Can be designed to create specific nucleotide substitutions, deletions, and so on
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Image of Site-directed Mutagenesis
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Two different forms of Site-specific Mutagenesis 5’ add-on mutagenesis - Diagram on page 148 of the text - a new sequence or chemical group is added to the 5’ end of a PCR product Mismatched primer mutagenesis - primer is designed to be only partially complementary
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Mismatched Mutagenesis Diagram pg. 147 of text; Figure 5.19 Can create a desired point mutation at a unique predetermined site within a cloned DNA molecule At the intended mutation site it bears a base that is complementary
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References http://humogef.ifj.edu.pl/zaklad/laborat or.html (pictures) Textbook http://www3.interscience.wiley.com/cg i- bin/jhome/10009058?CRETRY=1&S RETRY=0
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