L24 Detection and analysis of genes

L24 Detection and analysis of genes - Detection and...

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Detection and analysis of genes 1
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Last Time Making many copies of a gene Insertion of gene into a vector Making cDNA from RNA Restriction mapping PCR - polymerase chain reaction 2
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Hybridization based techniques: Detecting specifc clones in a DNA library (colony hybridization Detecting a specifc DNA Fragment ( Southern Blot ) Detecting a specifc RNA Fragment ( Northern Blot ) Protein-binding techniques: Detecting a specifc protein ( Western Blot ) Detection and analysis of genes 3
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Where we left off…. We have cloned a fragment of DNA ...but which colony contains our gene of interest?? 4
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Colony hybridization technique locating the clone that contains our gene of interest (don’t worry about colored colonies in this slide yet) Treat Flter to: dissolve cell membrane, denature DNA, bind DNA to Flter ±ilter Transfer some of the colony cells to a Flter 5
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Probing and detecting a gene sequence within a colony insulin gene colonies that contain insulin gene 6
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This note was uploaded on 03/29/2009 for the course BIO 325 taught by Professor Saxena during the Fall '08 term at University of Texas at Austin.

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L24 Detection and analysis of genes - Detection and...

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