[Yeast] RE: RT-PCR reference primers

Harrison, Eleanor via yeast%40net.bio.net (by e.harrison06 from imperial.ac.uk)
Wed Mar 28 04:13:53 EST 2007


Hi Lisa,

I've just started using primers based on STE2, which is a single copy
pheromone receptor gene, and it seems to be working really well.  The
only problem might be that, as it is a mating factor gene it is specific
to MATa cells. For me this is great as it is at single copy in diploids,
but may not be that helpful if you use haploid strains. I was also
suggested a couple more; STE3 (the MATalpha equivalent), AGA, SAG.  I am
not sure about NaCl stress but I'd imagine that the mating factor genes
would not be affected.

I hope this is helpful,

Ellie Harrison
Imperial College
UK

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Today's Topics:

   1. RT-PCR (Lisa Ooms)
   2. Re: RT-PCR (Maria Costanzo)


----------------------------------------------------------------------

Message: 1
Date: Tue, 27 Mar 2007 16:22:54 +1000
From: Lisa Ooms <lisa.ooms from med.monash.edu.au>
Subject: [Yeast] RT-PCR
To: yeast from magpie.bio.indiana.edu
Message-ID: <4608B83E.C843BCA from med.monash.edu.au>
Content-Type: text/plain; charset=us-ascii

I'm trying to find a reference gene for RT-PCR on S. cerevisiae
following hyperosmotic stress. Can anyone suggest a gene that does not
alter in response to NaCl stress?

Thanks for your help.

Lisa Ooms
Monash University
Australia



------------------------------

Message: 2
Date: Tue, 27 Mar 2007 10:33:36 -0400
From: Maria Costanzo <mcc from twcny.rr.com>
Subject: Re: [Yeast] RT-PCR
To: Lisa Ooms <lisa.ooms from med.monash.edu.au>
Cc: yeast from magpie.bio.indiana.edu,	SGD Curator
	<yeast-curator from genome.stanford.edu>
Message-ID: <ECB265C4-E8B4-40D4-B48C-E44574511306 from twcny.rr.com>
Content-Type: text/plain; charset="us-ascii"

Hi Lisa,

You may hear from someone who has used a particular gene for this  
purpose, but if not, you can use SGD's Expression Connection

http://db.yeastgenome.org/cgi-bin/expression/expressionConnection.pl

to investigate this. Using Search III, which allows you to find genes  
whose expression changes by the manner you specify, to find genes  
whose expression doesn't change significantly in the dataset  
"Expression in response to environmental changes" (A.P. Gasch et  
al.), should reveal some candidates. One of the conditions tested in  
this study was hyperosmotic stress.

Best regards,
Maria

Maria C. Costanzo, Ph.D.
Senior Scientific Curator, Saccharomyces Genome Database
   and Candida Genome Database
Department of Genetics
Stanford University School of Medicine
Stanford, CA 94305-5120
Phone: 650-725-8956
Fax: 650-723-7016
http://www.yeastgenome.org/
http://www.candidagenome.org/
maria from genome.stanford.edu


On Mar 27, 2007, at 2:22 AM, Lisa Ooms wrote:

> I'm trying to find a reference gene for RT-PCR on S. cerevisiae
> following hyperosmotic stress. Can anyone suggest a gene that does not
> alter in response to NaCl stress?
>
> Thanks for your help.
>
> Lisa Ooms
> Monash University
> Australia
>
> _______________________________________________
> Yeast mailing list
> Yeast from net.bio.net
> http://www.bio.net/biomail/listinfo/yeast


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