DNA from filter paper to use in transfomation - Molecular Biology (Aug/29/2005 )

Hi all--need some help again-
I have some DNA spotted on filter paper that I need to elute and use in a transformation. I plan to elute with TE (5-10 uL) and use about 0.5 uL in transformation. However, I have read some articles (from Whatman) that say that DNA eluted from filter paper to use in transfomation should be used for electocompetent cells, not chemically competent ones. (We use the latter, XL2Blue). Has anyone heard of this before?

Thanks,
slapolla

-slapolla-

QUOTE (slapolla @ Aug 29 2005, 01:07 PM)

I have done transformations into XL10 Gold cells in the last month from filter paper stored DNA. I added about 50 ul TE to the 5 ug DNA that was shipped to us, and it transformed just fine with 1 ul of the mix. So, it is probably worth the risk of 0.5 ul of your sample to get some colonies.

Good luck!

-cmp146-

Electroporation is more efficient than standard transformation so if you only have a small amount of intact DNA then you are more likely to get colonies from electroporation.

Daniel

Molecular biology help

-Daniel Tillett-

If you have supercompetent e.coli, than electoporation is really unnecessary. Remember, you realy only need one colony to grow your plasmid, right! BTW, I have done this numerous times from filter paper and never had a problem getting 20-50 colonies.

-tap14-

Thanks all! Will give it a try today.

slapolla

-slapolla-