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cloning upper stream of a gene


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#1 gee

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Posted 29 April 2004 - 11:00 PM

Dear all,

Hi, I am trying to clone a up stream of a gene. I cloned the partial gene and the problem is can't get the start codon (ATG) of the gene.

The gene I want to clone is rich in GC%. So for I have try to walk up using genomic as templet but failed. Is anyone can suggest what strategy can I use?

Thank you.

#2 pcrman

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Posted 30 April 2004 - 12:00 AM

Hi gee,

Please make it clear whether you want to clone the gene (full length mRNA) or its promoter.

I guess you want to clone the mRNA sequence. If that is the case, you have to do 5' RACE. You should not use genomic DNA as template, there may be introns which could be pretty big.

If you are interested in obtaining the promoter, do a genomic walk.




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