[Yeast] oleic acid medium

V McDonough via yeast%40net.bio.net (by mcdonough At hope.edu)
Mon Jan 15 13:11:45 EST 2007


Hi Anna-
I always prepare a 100X solution of my fatty acid in ethanol-you can filter 
sterilize if you think you must, but don't boil it.  I prepare my medium with 
1% tergitol NP-40 added to solubilize the fatty acid, than sterilize as usual.
 I add the fatty acid to my medium after it has been sterilized and cooled.  
Just keep in mind that you will have a final concentration of 1% ethanol in 
your medium.  For the work I do, I don't need potassium phosphate buffer in 
the medium, so I can't help you there.  
Virginia McDonough



>===== Original Message From "Anna Amelina" <aamelina At gmail.com> =====
>Hello again.
>Need your help! Does anyone can suggest me kind of standart protocol for
>preparing oleic acid medium, in particularly I'd like to use YPOT? I need to
>use it for peroxisomes proliferation induction in wild type S.cerevisiae.
>I've read a lot of articles and still can't find what I need. I've tried one
>from Thieringer, where they use 0,15%(w/v)oleic acid and 0,015%(v/v)Tween
>40. But I have some questions regarding:
>- potassium phosphate buffer: which molarity or precentage I need to use? In
>that article they suggest 30mM K2HPO4, and then to adjust pH of final medium
>to 6.0, but K2HPO4 is too basic.. As I understand I need it to get right pH
>only, so I tried to titrate with KH2PO4 to get pH 6. Is it correct way of
>doing it?
>- preparing oleic acid stock with Tween 40. Do I need to add them after
>autoclaving? What's the ratio for this components ( in different papers they
>use different). I did in a following way: made a stock of 150ul Tween 40 +
>1.5 g oleic acid, and boil it, then added to autoclaved medium, but I'm not
>sure that in this way olec acid willbe in solution. Ideally to get oleic
>acid into solution you need ethanol?
>Please, if you have your protocol for this or any suggestions, let me know!
>Thank  you in advance
>--
>Anna
>
>
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V McDonough
Associate Professor of Biology
Hope College
PO Box 9000
Holland MI 49422-9000
voice (616) 395-7715
fax (616) 395-7125



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