Gel Solubilization Buffer Composition

Suresh Kumar via methods%40net.bio.net (by kumar from lifesensors.com)
Mon Jan 11 16:13:39 EST 2010


Here is couple of different buffer composition.

Gel solubilization buffer -I 

60% Guanidine Thiocyanate (5.36M)
140mM MES (2-[N-Morpholino] ethanesulfonic acid)
0.006% Phenol Red
 	
Use 3 Volume of buffer to 1 gel volume.

Reference: http://www.plosone.org/article/info:doi%2F10.1371%2Fjournal.pone.0007750

Phenol Red is only a pH indicator. It changes color from yellow to red over the pH range 6.8 to 8.4.  

Gel solubilization buffer -II

6M Guanidine Thiocyante
50mM Tris-HCl pH 7.5
20mM EDTA pH 8.0

Use 3 Volume of buffer to 1 gel volume.


Hope this helps.	

Suresh

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Message: 2
Date: Sat, 9 Jan 2010 22:10:12 +0530
From: Jagadish Katam <jagadish.kv from hotmail.com>
Subject: Gel Solubilization Buffer Composition
To: <methods from magpie.bio.indiana.edu>
Message-ID: <SNT124-W189ABD0BA2F045D7F1CB96E76F0 from phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"


Hi,

I am working on purifying the DNA by gel extraction procedure and are using the commercially available kits for this purpose. A yellow color gel solubilization buffer is used to dissolve the sliced gel. 

Here i would know the composition of this gel solubilization buffer. 

Any suggestions will be greatly appreciated. 

Thanks and Best Regards,
Jag

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

Message: 3
Date: Sat, 9 Jan 2010 12:40:27 -0800 (PST)
From: Nick Theodorakis <nick.theodorakis from gmail.com>
Subject: Re: Gel Solubilization Buffer Composition
To: methods from net.bio.net
Message-ID:
	<f87be422-8f3e-4683-9cda-f1d3c01ac53e from j19g2000yqk.googlegroups.com>
Content-Type: text/plain; charset=ISO-8859-1

On Jan 9, 11:40 am, Jagadish Katam <jagadish... from hotmail.com> wrote:
> Hi,
>
> I am working on purifying the DNA by gel extraction procedure and are using the commercially available kits for this purpose. A yellow color gel solubilization buffer is used to dissolve the sliced gel.
>
> Here i would know the composition of this gel solubilization buffer.
>

In general, agarose gel solubilization can be done with chaotropic
salts such as NaI, sodium perchlorate, or guanidine HCl. These also
work well in conjunction with silica-based binding resins because DNA
binding to silica is promoted in high salt.  I doubt the company will
tell you exactly what is in their buffer, but it is likely similar to
one the salts I mentioned, possibly with additives added as a
stabilizer or other use.

Nick

--
Nick Theodorakis
nick_theodorakis from hotmail.com
contact form:
http://theodorakis.net/contact.html




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

Message: 4
Date: Sat, 9 Jan 2010 18:19:29 -0700 (MST)
From: "Dr. Hiranya S. Roychowdhury" <hroychow from nmsu.edu>
Subject: Re: hi .......help me plz
To: "Amol Ghodke" <amoltej from gmail.com>
Cc: methods from magpie.bio.indiana.edu
Message-ID: <1120.71.33.47.214.1263086369.squirrel from webmail.nmsu.edu>
Content-Type: text/plain;charset=iso-8859-1

Do you have access to a library? If you do, perhaps you may check out a
few of the Methods in Enzymology volumes for zymogram protocols.  If you
have access to the internet, Google search of the protocols (both zymogram
and protein elution from gels - whether acryl. or starch) will show you
several protocols.  Protein/peptide elution from gels is quite simple -
works with electrophoresis or simple diffusion/dialysis methods.


> hi all...
> i have to screen proteinase inhibitors!
> for that i have to do zymography.
> can any one plz give me the detail protocol for that
> and plz tell me how to recover screened protein from gel efficiently
> without
> using highly sofasticated instuments
> plz help me
> thanx
>
> --
> Mr. Amol Bharat Ghodke
> M.Sc. Agril.Biotechnology
> A/P - Kanhur Pathar
> Tal.-  Parner,
> Dist.- Ahmednagar
> Pin - 414303
> Mob. 09970865283/09021257663
> _______________________________________________
> Methods mailing list
> Methods from net.bio.net
> http://www.bio.net/biomail/listinfo/methods
>


-- 
Hiranya S. Roychowdhury, Ph.D.
Asst. Professor,
Health & Public Services
Dona Ana Community College
New Mexico State University
Las Cruces, NM 88003



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Message: 5
Date: Sun, 10 Jan 2010 11:06:34 +0200
From: Yoram Gerchman <gerchman from research.haifa.ac.il>
Subject: yeast estrogen screen?
To: methods from oat.bio.indiana.edu
Message-ID: <1263114394.4b49989acbeb4 from webmail.haifa.ac.il>
Content-Type: text/plain; charset=windows-1255


Greetings all
I am trying to get some version of the Yeast Estrogen Screen Assy for some
enviormental work (beta-gal, GFP, ...). Does anyone know were does one get the
yeast and plasmid from?
Many thanks
Yoram

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