Methods Digest, Vol 30, Issue 15

Virash Gupta via methods%40net.bio.net (by virashkgupta from gmail.com)
Fri Nov 23 13:12:18 EST 2007


For testing viability of culture, you need to explain the organism and
your experimental requirements. Different persons and similarly
different microbes act differently in different circumstances.
V K gupta
Prof Microbiology
Spec: Mol Biology

On Nov 23, 2007 10:34 PM,  <methods-request from oat.bio.indiana.edu> wrote:
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> Today's Topics:
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>   1. RE: Protein/protein-crosslinkers (Peter Henriksen)
>   2. RE:Protein/protein-crosslinkers (Ozan Aygun)
>   3. Re: bacterial viability assay (Pow Joshi)
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> ----------------------------------------------------------------------
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> Message: 1
> Date: Thu, 22 Nov 2007 18:17:44 +0100
> From: "Peter Henriksen" <peterh from my.molbio.ku.dk>
> Subject: RE: Protein/protein-crosslinkers
> To: <methods from magpie.bio.indiana.edu>
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>
> Thank you for your great answers.
>
> I realize that achieving the actual chemistry of crosslinking two carboxyl groups is a lot tougher than I anticipated.
>
> If I got it right, using a carbodiimide (like EDC) will be a problem for me because it prepares carboxyls to react with amines and thereby Proteins with DNA. Adding high concentrations of polyamines might out-compete DNA-amines but might also cause very little crosslinking since carboxyl groups have to bind the same polyamine-molecule to be crosslinked.
>
> The use of more target-specifik crosslinking methods (like transglutaminase under a controlled promoter) seems less attractive to me because I want the crosslinking to be fairly unspecifik and capable of crossbinding lots of different proteins.
>
> However, knowing that I have to rethink my strategy is also of great value.   :o)
>
> Cheers
>
> Peter Henriksen
>
>
>
>
> ------------------------------
>
> Message: 2
> Date: Thu, 22 Nov 2007 12:00:07 -0800 (PST)
> From: Ozan Aygun <metugenetics from yahoo.com>
> Subject: RE:Protein/protein-crosslinkers
> To: methods from magpie.bio.indiana.edu
> Message-ID: <608483.91501.qm from web90410.mail.mud.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> Hi Peter,
>
> Some years ago I have used a crosslinking reagent
> called as DSP. This should be available from Pierce.
> It is usually quite good when used properly, and thiol
> cleavable.
>
> I am not quite sure whether it fits your requirements
> but worth to try. There are even more crosslinkers
> offered by Pierce, perhaps you should look into their
> web catalog for properties.
>
> Best,
>
> Ozan
>
>
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> ------------------------------
>
> Message: 3
> Date: Thu, 22 Nov 2007 16:11:32 -0500
> From: "Pow Joshi" <pow.joshi from gmail.com>
> Subject: Re: bacterial viability assay
> To: "Sylvie Chauvaux" <chauvaux from pasteur.fr>
> Cc: methods from magpie.bio.indiana.edu
> Message-ID:
>        <710764ea0711221311o16b42312ocb7aa0e0952fa982 from mail.gmail.com>
> Content-Type: text/plain; charset=ISO-8859-1
>
> On 21/11/2007, Sylvie Chauvaux <chauvaux from pasteur.fr> wrote:
> >
> > Hello,
> >
> > Does somebody know an assay to follow bacterial viability of a
> > culture ? The commercial kits implicate to take samples of cultures.
> > Since I need to follow the viability overnight, I need an assay where
> > the reagents can be present in the culture without affecting growth
> > of bacteria.
> >
> > Many thanks for all your ideas,
>
>
>
>
>
> I don't claim to know too much about microbes, however would'nt plating the
> bugs either on regular  LB or mininal medum work? Ofcourse LB would need to
> have an antibiotic marker, I guess.
>
> Pow
>
> Sylvie
> >
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> End of Methods Digest, Vol 30, Issue 15
> ***************************************
>



-- 
Dr V K Gupta
Sr Microbiologist (Molecular Biology)
Insect Molecular Biology Lab
Department of Entomology
Punjab Agricultural University
Ludhiana (Pb)-141004- India
M: 09815963210



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