3_22 - DNA TECHNOLOGY Restriction endonucleases- cut within...

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DNA TECHNOLOGY Restriction endonucleases- cut within a DNA molecule (rather than digest it at the end) to make 2 pieces of DNA. These cut at a particular sequence that they recognize—this makes them great tools. See outline diagram. First letter- genus Next two letters- species Letter before roman numeral- strain EcoRI will break the phosphodiester bond between the guanosine and adenosine. But notice what’s on the other strand. It is dependent upon polarity… but DNA is antiparallel… the other strand reads exactly the same thing as the top strand, but backwards. This is called a restriction digest- breaking DNA into smaller fragments (restriction fragments). The single stranded overhangs are called sticky ends- the bases there could form hydrogen bonds if they could find a single-stranded complementary sequence to bind to. Take restriction digests from different digests and put them together to make a recombinant sequence…the sticky ends are just hydrogen bonds so they are momentary interactions…so… What would you need to reform these fragments into a stable molecule? Ligase, to reform the phosphodiester bonds. If you take restriction fragments and put
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This note was uploaded on 04/15/2008 for the course BIO 325 taught by Professor Saxena during the Spring '08 term at University of Texas at Austin.

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3_22 - DNA TECHNOLOGY Restriction endonucleases- cut within...

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