Colony PCR - (May/16/2011 )
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
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.