help in doing yeast transformations !!!

John S Jacobs Anderson jacobs at azstarnet.com
Mon May 5 08:19:48 EST 1997


Ed--

Check out the very informative (and not-at-all retentive) Yeast
Transformation Home Page at 

<http://www.umanitoba.ca/faculties/medicine/human_genetics/gietz/Trafo.html>

These guys (quite literally) wrote the book on yeast transformations...

John
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John S. J. Anderson                 http://www.azstarnet.com/~jacobs
Grad Student, MCB Dep¹t, U. of Az.       mailto:jacobs at azstarnet.com
My computer is so powerful that it does a infinite loop in 6 seconds!


In article <Pine.A41.3.95.970504174308.25840D-100000 at aix1.uottawa.ca>, ed
<s535290 at aix1.uottawa.ca> wrote:

>Hi all,
>
>Our electroporator is on the fritz, and I need to do yeast transformations
>preferably using the LiAc method. I have a somewhat modified (and
>much simpler !!!) version from the one that appears on the red book.
>Nonetheless, I read the red book for pointers...the thing that struck me
>was that it mentions that single most critical parameter in transformation
>efficiency when using the LiAc method is the quality of the carrier DNA
>being used...in particular, that it be totally denatured and of a median
>fragment size of about 7 Kb (!?!?!?). The question I ask of anyone who's
>done yeast transformations, is whether you prepare your own carrier DNA or
>purchase it from somewhere ?  It mentions sonication in the shearing of
>the DNA, but without access to a sonicator, I don't know how to go about
>getting the fragment sizes right...in the red book they even suggest
>running an aliquot to check median fragment size...are they being
>unnecessarily anal retentive about this or should I definitely be
>concerned about this ? I don't need an incredible transformation
>efficiency, as long as I have a few different transformants...please share
>your knowledge with me :)
>
>thanks in advance,
>
>Ed



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