Lecture25S11 - BIS101/Engebrecht Lecture25 5/27/11...

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BIS101/Engebrecht Lecture25 5/27/11 Recombinant DNA molecules are sequences that result from molecular cloning to bring together genetic material from multiple sources. Molecular cloning refers to a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms. Cloning refers to the fact that the method involves the replication of a single DNA molecule starting from a single living cell to generate a large population of cells containing identical DNA molecules. Genomic libraries : These are clone banks that contain genomic DNA inserts that cover the entire genome. Obviously, the larger the insert size, the small number of different clones will be needed to cover the entire genome. How do you identify your clone of interest from a DNA library? We discussed two approaches: 1) DNA homology : If you know some sequences of the gene you are interested in, you can use the properties of Watson-Crick base pairing and the principles of denaturation/renaturation. PCR is an example of this (we will talk about this on Tuesday). To find your DNA of interest in this case, you would need to have a library of E. coli clones. You would then make a replica of the E. coli colonies harboring different recombinant plasmids on a filter. The cells on the filter are lysed, the DNA bound to the
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Lecture25S11 - BIS101/Engebrecht Lecture25 5/27/11...

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