11-DNA cloning I-10

11-DNA cloning I-10 - DNA cloning I: Fragmenting genomes...

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DNA cloning I: Fragmenting genomes into smaller pieces and cloning them into plasmid vectors 11/15/10
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What does it mean to clone DNA? Steps in DNA cloning 1. Cleave long DNA molecules into smaller pieces. 2. Ligate individual DNA fragments to a cloning vector. 3. Transform recombinant DNA into bacteria. 4. Screen to identify clones of interest. 5. Prepare large amounts (many copies ) of a particular DNA clone to use in experiments.
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Restriction site in DNA, showing symmetry of the sequence around the center point
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Making recombinant DNA using the restriction enzyme Eco RI
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  Restriction fragments can be inserted into a plasmid vector so that the fragment can be replicated in E. coli
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Restriction enzymes Fig. 9.2
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Fig. 9.3.a
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Fig. 9.3.b
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Partial digests of DNA with a restriction enzyme Fig. 9.4.a
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Gel electrophoresis separates DNA molecules according to size. Fig. 9.5.a
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Fig. 9.4 When genomic DNA is digested with a restriction enzyme for increasing amounts of time, the DNA is digested more completely, resulting in shorter fragments.
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This note was uploaded on 02/24/2011 for the course BIOL 202 taught by Professor Kieber-hogan during the Fall '08 term at UNC.

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11-DNA cloning I-10 - DNA cloning I: Fragmenting genomes...

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