Long range Genomic PCR

[Dave Adelson]

In message <376nij$2bu at netnews.upenn.edu> - jlanders at hmivax.humgen.upenn.edu. (
jlanders) writes:
>I am interested in finding the genomic region of a cDNA that I have in
>my possession.  I am thinking about attempting to use long range PCR
>using primers from the cDNA with genomic DNA.  I also would like to
>obtain the promoter region and thought that maybe I could use a gene
>specific primer with a vector specific primer and perform longe range
>PCR using DNA derived from a cosmid library as my template.  Has anyone
>tried this or have any suggestions? I would appreciate any comments.


I can't help you regarding the long range PCR issue, but I have tried PCR 
cloning of a promoter region with no success.  My promoter had inverted 
repeats in it (not uncommon really) and a high GC content (not uncommon 
either) and it would not PCR.  We wound up subcloning it from a genomic clone 
directly after wasting 2.5 months on PCR in it's infinite and seductive 
variations.

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*Dave Adelson  "why ask me?" *
*CSIRO Div. Animal Production*
*Sydney, Australia           *
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