XbaI and genomic DNA digestion

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Gauch01
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XbaI and genomic DNA digestion

Post by Gauch01 » Thu Jan 25, 2007 3:38 pm

I have difficulty to digest genomic DNA from mouse tail using the XbaI restriction enzyme. Although, the enzyme work perfectly with genomic DNA from ES cell I cannot perform a good southern using the mouse tail genomic DNA. Others enzymes such EcoRI and BamHI work very well but not XbaI. I had concern about Quality of the DNA so tried different methods of purification ( phenol/chloro; quiagen column, spooling techniques), none of them work. I also tried XbaI enzyme from different suppliers and to heat inactivate the inhibtor factor that could be present in the DNA, without success either.

Any suggestion will be welcome!

Christian

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canalon
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Post by canalon » Thu Jan 25, 2007 6:27 pm

According to this site:
http://rebase.neb.com/rebase/enz/XbaI.html
XbaI is methylation sensitive. I am not familiar with mouse methylation systems but it might be a cause of your problem? Otherwise I don't have any idea.
Patrick

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)

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