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Cloning a promoter region [View Printable]
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parvoman
Group: Member Posts: 190 Joined: Jul 28, 2005
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I'm thinking about doing some analysis of a gene's enhancer / promoter region and was wondering how best to go about it.
Previously I would have looked for a human genomic library, done Southern using a probe to the coding sequence (lysyl oxidase - also known as LOX) to pick a bacterial colony containing the plasmid with the LOX fragment of interest. Then sequenced and performed PCR to isolate the 2-3Kb region of interest which would then be cloned upstream of a reporter gene in another plasmid to allow analysis of enhancer / promoter activity.
Is this still the way to do it or am I a dinosaur?
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| Posted Jan 19, 2008, 1:02 AM |
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parvoman
Group: Member Posts: 190 Joined: Jul 28, 2005
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Have found out that there is a BAC clone for this. This should make it easier.
Comments, any experience out there?
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| Posted Feb 08, 2008, 3:56 AM |
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