dna_restriction_maps

dna_restriction_maps - Biocomputing University of North...

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Biocomputing University of North Texas Biocomputing University of North Texas 1 University of North Texas Biocomputing DNA Restriction Maps Source: www.bioalgorithms.info Biocomputing University of North Texas Molecular Scissors Molecular Cel Biology, 4 th edition Biocomputing University of North Texas Discovering Restriction Enzymes Hin dII - first restriction enzyme – was discovered accidentally in 1970 while studying how the bacterium Haemophilus influenzae takes up DNA from the virus Recognizes and cuts DNA at sequences: GTGCAC GTTAAC Biocomputing University of North Texas Recognition Sites of Restriction Enzymes Molecular Cel Biology, 4 th edition Biocomputing University of North Texas Restriction Maps A map showing positions of restriction sites in a DNA sequence If DNA sequence is known then construction of restriction map is a trivial exercise In early days of molecular biology DNA sequences were often unknown Biologists had to solve the problem of constructing restriction maps without knowing DNA sequences Biocomputing University of North Texas Full Restriction Digest Cutting DNA at each restriction site creates multiple restriction fragments : Is it possible to reconstruct the order of the fragments from the sizes of the fragments {3,5,5,9} ?
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Biocomputing University of North Texas Biocomputing University of North Texas 2 Biocomputing University of North Texas Full Restriction Digest: Multiple Solutions Alternative ordering of restriction fragments: vs Biocomputing University of North Texas Measuring Length of Restriction Fragments Restriction enzymes break DNA into restriction fragments. Gel electrophoresis is a process for separating DNA by size and measuring sizes of restriction fragments Gel electrophoresis can separate DNA fragments that differ in length in only 1 nucleotide for fragments up to 500 nucleotides long Biocomputing University of North Texas Gel Electrophoresis: Example Direction of DNA movement Smaller fragments travel farther Molecular Cel Biology, 4 th edition http://www.dnalc.org/ddnalc/resources/electrophoresis.html Biocomputing University of North Texas Partial Restriction Digest The sample of DNA is exposed to the restriction enzyme for only a limited amount of time to prevent it from being cut at all restriction sites This experiment generates the set of all possible restriction fragments between every two (not necessarily consecutive) cuts This set of fragment sizes is used to determine the positions of the restriction sites in the DNA sequence Biocomputing University of North Texas Partial Digest Example
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This note was uploaded on 04/15/2011 for the course BIOL 1130 taught by Professor Roberts during the Spring '08 term at North Texas.

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dna_restriction_maps - Biocomputing University of North...

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