Methods Digest, Vol 17, Issue 23

Sayyari, Mohammad via methods%40net.bio.net (by msayyar from gwdg.de)
Wed Oct 25 04:11:58 EST 2006


Dear Sir/Madam
 
I am trying to measure oxidizer -reducer microorganism in rhizosphere soil.
Can one help me and tell the method? I would appreciate it, if you could help
me.
 
Best regards,
Mohammad

  _____  

From: methods-bounces from oat.bio.indiana.edu on behalf of
methods-request from oat.bio.indiana.edu
Sent: Tue 10/24/2006 7:02 PM
To: methods from magpie.bio.indiana.edu
Subject: Methods Digest, Vol 17, Issue 23



Send Methods mailing list submissions to
        methods from net.bio.net

To subscribe or unsubscribe via the World Wide Web, visit
        http://www.bio.net/biomail/listinfo/methods
or, via email, send a message with subject or body 'help' to
        methods-request from net.bio.net

You can reach the person managing the list at
        methods-owner from net.bio.net

When replying, please edit your Subject line so it is more specific
than "Re: Contents of Methods digest..."


Today's Topics:

   1. Re: low molarity borate and acetate agarose gels (Pow Joshi)
   2. Re: Stripping Solution (Pow Joshi)
   3. Wessel-Flugge Method for Protein Precipitation (Jeremy Kamil)
   4. pEGAD,pGreenII and p35S-GFP-JFH1 (Jinxiang Wang)


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

Message: 1
Date: Mon, 23 Oct 2006 15:41:49 -0500
From: "Pow Joshi" <pow.joshi from gmail.com>
Subject: Re: low molarity borate and acetate agarose gels
To: "Ho-Leung Ng" <holeung from berkeley.edu>
Cc: Methods from magpie.bio.indiana.edu
Message-ID:
        <710764ea0610231341g7406872egc17c77adbc37d2a4 from mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed

Hi,

frankly, I had run, regularly, some TBE gels (tris borate EDTA) and
the protocol is available in the Molecular cloning book, especially
the old editions by Tom Maniatis.....
You could use 0.5X instead of 1x buffer; alternatively, you could use
low voltages.
I used to find the resolutions comparable .... although it's easy to
run TAE gels if you wish to isolate/ purify your DNA.

hope this helps

pow
On 10/19/06, Ho-Leung Ng <holeung from berkeley.edu> wrote:
>      What are people's experiences using the Brody and Kern (Biotechniques
> 36:214 and 37:598) low molarity sodium borate and lithium acetate
> agarose gels? They do run very fast with little heat, but we often
> get very smeared bands, with borate performing better than acetate.
> There is a lot of band quality variability from run to run. Any tips
> would be appreciated!
>
>
> ho
> UC Berkeley
>
> _______________________________________________
> Methods mailing list
> Methods from net.bio.net
> http://www.bio.net/biomail/listinfo/methods
>



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

Message: 2
Date: Mon, 23 Oct 2006 16:20:20 -0500
From: "Pow Joshi" <pow.joshi from gmail.com>
Subject: Re: Stripping Solution
To: "Christian Praetorius" <prae from gmx.net>
Cc: Methods from magpie.bio.indiana.edu
Message-ID:
        <710764ea0610231420i24c17122y2e75e60b1e9674f0 from mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed

On 10/15/06, Christian Praetorius <prae from gmx.net> wrote:
> Maryam <mzargh from yahoo.ca> wrote:
>
> >Does anybody know if NaOH solution can be used as a stripping solution? if
> So, what should be the final concentration?

you might want to add some beta mercaptoethanol or DTT as well ...
this would cleave the -S-S- linkages in the antibodies (again assuming
you want to strip western blots)

pow
>
> What do you want to strip?
>
> Christian
>
> --
> X-no-Sig: yes
> _______________________________________________
> Methods mailing list
> Methods from net.bio.net
> http://www.bio.net/biomail/listinfo/methods
>



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

Message: 3
Date: Tue, 24 Oct 2006 02:29:19 -0400
From: Jeremy Kamil <jeremy.kamil from gmail.com>
Subject: Wessel-Flugge Method for Protein Precipitation
To: methods from magpie.bio.indiana.edu
Message-ID: <E4DEA952-3A7A-45B8-B8D8-F9AF2114DC17 from gmail.com>
Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed

Hi, I'm trying to concentrate dilute protein mixtures in my tandem 
affinity purification eluates for mass spec analysis. Specifically, I 
need to get rid of the EDTA and EGTA as well as the 0.1% tween or 
NP-40 that is in my elution buffer.

I saw that some people use the Wessel-Flugge method (Anal Biochem 
1974) for this application (Methanol, Chloroform precipitation). 
However, in my hands I found it didn't seem to work great, at least I 
don't see a nice pellet.

I've had some success in the past with TCA but often end up with some 
residual acidity even after many acetone washes. Anyway, at least 
with TCA I got a nice pellet every time.

My question re: Wessel-Flugge is, HOW MUCH OF THE UPPER LAYER ARE YOU 
SUPPOSED TO REMOVE??? Should I get rid of the upper phase all the way 
down to the chloroform layer, or leave a bit above of the chloroform??

The original publication says to remove the upper layer above the 
chloroform and interphase. But I don't see much of an interphase, 
except in some samples where I saw a diffuse band of white above the 
chloroform layer on the bottom. so, in case that diffuse white stuff 
was my protein, I left maybe 150 uL on top of the chloroform. In the 
end I did not see much, if any, precipitate and was pretty 
disappointed since the method seemed so promising.

any advice would be very much appreciated. I found the description by 
Wessel and Flugge pretty cursory.

thanks,

Jeremy





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

Message: 4
Date: Tue, 24 Oct 2006 19:39:49 +0800
From: Jinxiang Wang <jinxwang from scau.edu.cn>
Subject: pEGAD,pGreenII and p35S-GFP-JFH1
To: arab-gen from magpie.bio.indiana.edu, methods from magpie.bio.indiana.edu
Message-ID: <453DFB85.40303 from scau.edu.cn>
Content-Type: text/plain; charset=GB2312

Hello,All,

I appreciate someone who would like to share pEGAD,pGreenII and
p35S-GFP-JFH1 with me to construct GFP-tagged plasmid to transform
/Arabidopsis/. Thank you in advance.

Jinxiang

--
Jinxiang Wang Ph.D
College of Resources and Environment
Root Biology Center
South China Agricultural Univeristy
Guangzhou 510642, P.R.China
Tel:0086-20-85280156
Fax:0086-20-85281829
Email:jinxwang from scau.edu.cn



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

_______________________________________________
Methods mailing list
Methods from net.bio.net
http://www.bio.net/biomail/listinfo/methods

End of Methods Digest, Vol 17, Issue 23
***************************************





More information about the Methods mailing list