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Colony PCR


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#1 AGM1

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Posted 16 May 2011 - 06:46 PM

Hi,

I have created a genomic library that contains environmental insert DNA, and the host is E.coli. The library is made of fosmids, and the copy number is kept at one. I was planning on using colony pcr to screen the inserts present in the library. I was wondering if its possible to use
gram-positive pcr primers to screen the library, so as to avoid the host E.coli DNA, and still be able to screen for a large number of bacterial 16S genes. If anyone with expierience in these techniques could shed some light, it would be appreciated

Thank you

#2 BioMiha

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Posted 17 May 2011 - 09:01 PM

Why don't you use fosmid specific primers. That's how it is usually done. That way you don't need to worry about amplifying the host gDNA.




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