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[Yeast] Acid-washed glass Beads

David Mueller via yeast%40net.bio.net (by david.mueller from rosalindfranklin.edu)
Tue May 27 16:27:22 EST 2008


Cinzia:

We do it all the time - but we are not isolating DNA.


WASHING Glass Beads
 
Add beads to a 4 liter beaker with detergent and let sit overnight.
 
Rinse well with house distilled water.

Wash with 1M nitric acid overnight. (A 70% solution of Nitric acid is
15.4M.) 

Rinse with glass distilled water.

Dry in oven.  Beads should not clump.  If they do, they are not clean.

Beads can be reused up to 10 times.

David Mueller


On 5/27/08 11:18 AM, "Cinzia Pagliuca" <cinthiait from yahoo.com> wrote:

> Dear all,
> 
> I' m triyng to break the yeast cells with a Bead-Beater. Since I use 100-200
> ml of Glass Beads (sigma, acid washed Beads), I would like to re-use them. It
> is possible? Can you suggest how to do it?
> 
> Many thanks in advice,
> Cinzia
> 
> 
> --- On Mon, 5/26/08, yeast-request from oat.bio.indiana.edu
> <yeast-request from oat.bio.indiana.edu> wrote:
> 
>> From: yeast-request from oat.bio.indiana.edu <yeast-request from oat.bio.indiana.edu>
>> Subject: Yeast Digest, Vol 36, Issue 13
>> To: yeast from magpie.bio.indiana.edu
>> Date: Monday, May 26, 2008, 7:04 PM
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>> Today's Topics:
>> 
>>    1. Re: Raffinose versus Sucrose (Ilya Soifer)
>> 
>> 
>> ----------------------------------------------------------------------
>> 
>> Message: 1
>> Date: Mon, 26 May 2008 10:53:11 +0300
>> From: "Ilya Soifer" <ilya.soifer from gmail.com>
>> Subject: Re: [Yeast] Raffinose versus Sucrose
>> To: yeast from magpie.bio.indiana.edu
>> Message-ID:
>> <1952757a0805260053h60b91c14x30299960d15dd95c from mail.gmail.com>
>> Content-Type: text/plain; charset="iso-8859-1"
>> 
>> Dear all,
>> I have a similar question - some labs use autoclaved
>> raffinose and some -
>> filtered raffinose in the media prior to galactose
>> induction.
>> Does anyone have an idea why and which way is preferrable?
>> Thanks a lot,
>> Ilya
>> 
>> 2008/5/24 Daniel Bosch <daniel.bosch from gu.se>:
>> 
>>> Hi Kerry,
>>> 
>>> Both sucrose and raffinose are broken up outside of
>> the cell by the same
>>> enzyme, invertase. Sucrose generates glucose and
>> fructose that can be
>>> immediately transported into the cell and assimilated.
>> Raffinose leads to
>>> fructose and melibiose. The latter is broken up by
>> another enzyme,
>>> melibiase, which is not present in all lab strains. As
>> a matter or fact, the
>>> degradation of raffinose will yield less amount of
>> fermentable sugars (33%)
>>> in lab strains than sucrose (100%) and therefore it
>> can be considered a
>>> poorer carbon source. Neither sucrose nor raffinose
>> should lead to a major
>>> repression of the GAL genes.
>>> Having said this, I don't know why some labs
>> prefer sucrose over raffinose
>>> or mixtures of both. An alternative to sugars is the
>> use of respiratory
>>> carbon sources, such as glycerol or ethanol. I
>> particular find that cells
>>> grow better in a mixture of the two (3% glycerol and
>> 1% ethanol) than on any
>>> of them alone.
>>> 
>>> Hope this was helpful,
>>> 
>>> Dani
>>> 
>>> 
>>> Daniel Bosch
>>> Postdoc in Nyström's lab
>>> Dept. Cell and Molecular Biology
>>> University of Gothenburg
>>> Sweden
>>> 
>>> 
>>> ------------------------------
>>> From Kerry Geiler <kgeiler from gmail.com>
>>> Sent Sat 5/24/2008 12:50 AM
>>> To yeast from magpie.bio.indiana.edu
>>> Cc
>>> Subject [Yeast] Raffinose versus Sucrose
>>> 
>>> HI,
>>> I have a question about whether yeast grow better in
>> sucrose and raffinose
>>> or a combination of both.  I am using galactose
>> induction, so I cannot grow
>>> my strains in glucose.  I have inquired with many
>> nearby labs and some use
>>> 2% sucrose, some use 2% raffinose, and some use 2%
>> sucrose, 1% raffinose.
>>>  However, none of these labs can tell me why they made
>> this decision (it
>>> seems to be a part of their lab culture).  Does anyone
>> know the differences
>>> between yeast growth in sucrose vs raffinose?  Do
>> either of these sugars
>>> inhibit metabolism of the other sugar?  Any
>> information that will inform my
>>> decision of which sugar to choose would be greatly
>> appreciated.  Thanks!
>>> 
>>> Kerry Geiler
>>> kgeiler from gmail.com
>>> ------------------------------
>>> 
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David Mueller
Biochemistry and Molecular Biology
Rosalind Franklin University of Medicine and Science
3333 Greenbay Road
North Chicago, IL 60064
David.mueller from rosalindfranklin.edu
http://web.mac.com/mueller.david/iWeb/LMB/dm.html
Tel: 847-578-8606
FAX: 847-578-3240





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