TSS Transformation problems

Chung Jui Tsai chtsai at MTU.EDU
Thu May 29 16:36:00 EST 1997


Jared,

When I first saw the protocol, I also liked the 2X TSS one better, simply
because it's super easy -- all I need is 1:1 dilution and the cells are
ready to go!!
I filter sterilized LB+MgCl2+PEG then added DMSO and it was only a bit
cloudy as I remember.  However, I did not have any luck in making good
competent cells with 2X TSS (actually I tried only once) -- I did not get
any transformants using these cells in a test run (with other competent
cells).  Then I tried the 1X TSS protocol with one extra centrifugation
step, and IT WORKS GREAT.  In fact, my 1X TSS slou has been sitting at 4C
for more than a year and is still working fine......  

Hope this helps.

Tsai

At 10:30 AM 5/28/97 GMT, you wrote:
>Hi,
>
>I'm having a go at the quick and easy transformation method of Chung et
>al. as advertised in this newsgroup, but I'm concerned about making up the
>initial 2x TSS solution.  When I mix 20% PEG 4000, 10% DMSO, and 100mM
>MgCl2 with my 2xLB I get a very murky looking mixture, not all of which is
>dissolved.  Is this what you expect, and how am I supposed to sterilise
>it?  Filter sterilisation gives a nice clear liquid, but I suspect it's
>taken out some of the ingredients. 
>
>Thanks in advance for any help,
>
>Jared
>
>-- 
>Jared Head     at the Department of Biochemistry, University of Bristol
>
>"A computer lets you make more mistakes faster than any invention in human
>     history - with the possible exceptions of handguns and tequila." 
>                                                   Mitch Ratliffe
>
>




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