TSS Transformation problems

Joe Boutell boutell at cf.ac.uk
Thu May 29 03:47:50 EST 1997


bijgh at zeus.bris.ac.uk (Jared Head) 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

Hi Jared!
When we make up TSS solution we make it up to 1X only, and then autoclave 
to sterilize. I guess heating it in the autoclave will dissolve the 
cloudy stuff. We also make it fresh each time we make a batch of cells.
Cheers,
Joe Boutell.




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