Co transformation frequencies

Douglas Rhoads drhoads at MERCURY.UARK.EDU
Sat Nov 2 09:30:02 EST 1996


> To:            yeast at net.bio.net
> From:          Kathy Baynton <baynton at esbs1.u-strasbg.fr>
> Subject:       Question and email for list
> Date:          2 Nov 1996 11:26:35 GMT
> To:            francis at ncbi.nlm.nih.gov

> Dear Francis, 
> 
> 
> I am currently using electroporation to transfect my yeast with the 
> desired plasmid and was wondering if anyone knew what might be the level 
> of cotransformation using such a technique.  Most people I've talked 
> with suggest that this technique has a lower level of cotransformation 
> than either the lithium chloride, PEG or spheroplasting techniques.  But 
> some tests I've performed which enables the detection of two different 
> plasmids present in a single yeast colony arising after transformation 
> with an equal mixture of the two plasmids suggests that the level of 
> cotransformation can vary from 1-10%.  Carrier DNA is not used and is 
> usually not included in such a procedure.  I was wondering if anyone 
> knows any more about cotransformation levels using any of the common (or 
> not so common) transfection techniques and if possible, how can this be 
> minimized, IF at all possible. ANY suggestions, comments etc. would be 
> really appreciated!!  Thanks!!
> 
> Kathy Baynton

I too would be interested to know whether anyone has compared 
electroporation to PEG or LiCl transformation for frequency of 
co-transformation.  
Kathy, I  hope if you get helpful responses you will post a summary 
back to the group.
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||                  Douglas Rhoads                        ||                             ||
||drhoads at mercury.uark.edu || Dept. of Biological Sciences||
||drhoads at comp.uark.edu    || 601 Science Engineering     ||
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