please help

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ooolex
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please help

Post by ooolex » Sun Feb 05, 2006 5:46 pm

Perhaps someone here can help me with a few questions concerning a housework.

The task is the elaboration of a procedure way for the insertion of a marker suitable for recombination mapping in S.cerevisiae. It is to be inserted between ADE5,7 and LYS5 at the chromosom VII.

Now my idea would have been to manage this by use of a disruption-cassette which contains short homologous flanking sequences of the gene to disrupt and in between the marker gene, I thought of NeoR or KanR.
For the gene to disrupt every gene is suitable that produces a viable phenotype, correct?

Further questions emerged by the fact that the Professor explained a problem similar to this but he used a Yeast integration Plasmid instead of a disruption cassette. The YIP contained the marker and the gene, which is in the desired range. Then the Plasmid is to integrate by homologous recombination at the desired place including the marker.

now I don't know whether both possibilities would work or whether there are reasons against one possibilty.

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