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methylated plasmid - (Jun/08/2006 )

I am trying to transfect a bacterium that is sensitive to methylation but my plasmids are from a bacterium that methylates. Can I transfect an E. coli that doesn't methylate plasmids and receive unmethylated plasmids?

-catrina-

many common used E.coli sgtrains have been modified not to have DNA methylase,such as DH5a,Top10,you can just try!Good luck!

QUOTE (catrina @ Jun 9 2006, 08:59 AM)
I am trying to transfect a bacterium that is sensitive to methylation but my plasmids are from a bacterium that methylates. Can I transfect an E. coli that doesn't methylate plasmids and receive unmethylated plasmids?

-JerryLee-

Check out stratagene, invitrogen, neb for e. coli that doesn't methylate, most strains (the mentioned DH5a, Top10) do methylate, but there are some dam- and dcm- ones around.

links (probably other companies supply them as well):
http://www.neb.com/nebecomm/products/productC2925.asp
http://www.stratagene.com/products/display...ct.aspx?pid=303
http://www.invitrogen.com/content.cfm?page...44#preparing_un

-vairus-

Thanks for the help. I have found one from New England Biolabs smile.gif

-catrina-