to S.L. Rennie -trans prot

Kathy Shire shirek at FHS.CSU.MCMASTER.CA
Fri Nov 4 15:45:13 EST 1994





---------- Forwarded message ----------
Date: Wed, 02 Nov 1994 15:11:34 -0600
From: "R. D. Gietz" <GIETZ at BLDGHSC.Lan1.umanitoba>
To: yeast at net.bio.net
Subject: Re: two plasmids in yeast

In article <38uiol$pij at newsbf01.news.aol.com>, brianc4208 at aol.com
(BrianC4208) wrote:

> I agree that it is easier to transform with both plasmids at once, but I
> didn't realize that you could transform two plasmids with 2 micron
> replication origins into the same cell.  I was always under the impression
> that you had to have one on a centromeric plasmid.  Please correct me if I
> am mistaken, because it would make a big difference in an experiment I'm
> currently setting up for this week.
> 
> Thanks,
> Brian Cohen
 > Albany Medical College
> BrianC4208 at aol.com


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Hey there
   I don't know if you EP or use LiAc to trafo, but if you use LiAc, I
know for a fact that you can stick in two at a time, I think we even did
three at once but then the efficiency is way down.

My boss has a bunch of good trafo papers out:
   Shiestl et. al. "Introducing DNA into yeast by Transformation" in
Methods: A Companion to methods in Enzymology.

I can't find the more recent paper in Methods in enz 'cause we're in the
midst of a lab move.  But I find that our methods are quicker easier and
less full of supersitions (though there are still some left in ??) than
other things I've heard people doing. 

Anyway save some time, live on the edge and definately do both at once, 
best to check copy number of each so you don't swamp with one.

-- 
S.L.Rennie
Department of Human Genetics
University of Manitoba
Winnipeg, Manitoba, CANADA
GIETZ at BLDGHSC.LAN1.UMANITOBA.CA

Hi S.L.!

If you can find some time between moving valuable equipment around, could
you send me some more info on these protocols?  I would like to know if my
protocols are filled with myths (there is no mention of Hercules and his
magic ring, so they can't be that bad).  date/pg/vol# would be greatly
appreciated.

Thanks

Kath






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