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suggestions for new PAGE apparatus

suggestions for new PAGE apparatus - Protocols and Methods Forum

suggestions for new PAGE apparatus - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 08-03-2005, 01:52 PM
Dr. James J. Campanella
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Default suggestions for new PAGE apparatus



Hi everyone,

I am in the market to buy a new PAGE set-up. I refuse to buy Biorad again.
I have been burnt by their low quality equipment before. We have begun
pouring a large number of gels, and I am sick of wasting half my time time
trying to keep the casting system from leaking even as I'm pouring my gel.
There has to be something better out there than Biorad. I would like
recommendations on which company's apparatus will allow me to pour my PAGEs
every time without leakage.

Thanks for the help,

Jim

James J. Campanella,
Associate Professor
Department of Biology and Molecular Biology
Montclair State University
1 Normal Avenue
Montclair, NJ 07043

Alternate email address: [Only registered users see links. ]

Ph: 973-655-4097
Fax: 419-791-9834

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  #2  
Old 08-04-2005, 12:07 AM
Bean Long
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Default suggestions for new PAGE apparatus

It may not suit but I would suggest precast gels. Saves heaps on time and
grief. We use Invitrogen precast NuPAGE gels and run them in an old Hoefer
rig which uses very little buffer. No complaints after close to 100 gels so
far.

I know that BioRad also have their Criterion range of pre-casts and we have
one of their rigs but the gels are to big for our use. The have something
like a minimum of 18 lanes (I could be wrong here) and we can't find the
samples to fill them. Nonetheless, they would certainly suit particular
applications.

Best of luck,
--
Bean

Remove "yourfinger" before replying

"Dr. James J. Campanella" <[Only registered users see links. ]> wrote in message
news:[Only registered users see links. ]. net...
time
PAGEs


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  #3  
Old 08-04-2005, 03:59 AM
Nick Theodorakis
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Default suggestions for new PAGE apparatus

On Wed, 03 Aug 2005 09:52:22 -0400, "Dr. James J. Campanella"
<[Only registered users see links. ]> wrote:


I don't have a good suggestion for a replacement, but what I often
used to do with the Biorad setup is to put a strip of Parafilm between
the plates and the gasket. Alternatively, if I think the plates are
really dinged up, I make a stock of 1% agarose in the same Tris-Cl
buffer the gels are made with; then heat it up and put a bead of
agarose down each edge of the plates and the bottom. The agarose trick
dates back from the "home-made"gel plates days and is usually
fool-proof.

Nick

--
Nick Theodorakis
[Only registered users see links. ]
contact form:
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  #4  
Old 08-04-2005, 03:26 PM
Chris B.
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Default suggestions for new PAGE apparatus

> I am in the market to buy a new PAGE set-up. I refuse to buy Biorad again.

I use Sigma Mini Vertical - cat.no. Z339571 - and I'm quite happy with it.
You can also buy module for 2D electrophoresis for this one.

time
PAGEs

Usually to prevent leakage I prepare small quantity of gel with more TEMED
and APS then needed. I use it quickly to make "seal" on bottom and sides of
plate (1-2 mm).


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  #5  
Old 08-04-2005, 03:45 PM
Rafael Maldonado
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Default suggestions for new PAGE apparatus

Jim, we use mini-gel set-up from Hoeffer-Pharmacia-Amersham and I like
them most. They don't leak, are easy to clean and they sell replacements
for everything that can be wore out. No relation with the company,
except for decades using their products through different company names.

-R



Dr. James J. Campanella wrote:

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  #6  
Old 08-04-2005, 05:13 PM
IanMc
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Default suggestions for new PAGE apparatus

Hi,

The easiest system I ever used was from Atto, basically it was a vertical
gel tank, two nice thick glass plates with the spacers glued onto one side
and a gasket which went round the outside to form a U. It never leaked, was
difficult to break and took about 20s to set up. If you do go down this
route make sure you buy the first version of the system not the one with the
flappy hands. It is bomb-proof.

Ian Mc

"Dr. James J. Campanella" <[Only registered users see links. ]> wrote in message
news:[Only registered users see links. ]. net...


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  #7  
Old 08-04-2005, 11:01 PM
kaj.stenberg@helsinki.fi.invalid
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Default suggestions for new PAGE apparatus

Nick Theodorakis <[Only registered users see links. ]> wrote:


(...)


Tape (the firm and sticky type) does the trick even easier and faster.
Never mind any glue-residues, you have a reference there.

I have this problem in a course, some students are making their first
gels ever. Some succeed immidiately, some make unsuccesful trials 3
times and in the end have to be given mymade gels. And yes, it is
the BioR-apparatus.
Mishap, mistakes, failure and frustration are important learning
points in any wet-lab course. However, often there will be something more
intended than the mere basics. Basics should vork at the student's
level, I am sick and tired of telling frustrated students (in effect)
that "it works for me, and it is commersial, therefore it must be your
shortcomming, take this gel and continue your work".

Hovewer, since it is a marketed product (Bio-Rad apparatus) there must
be someone at Bio-rad who can cast these gels manually and routinely
without leaking problems and who cannot cannot see the problems we
meet in our first 100 gels. Of course, one can pay for the precasted
ones, is that the (failed) long-term plan?

--
Kaj
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  #8  
Old 08-09-2005, 08:31 AM
MMu
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Default suggestions for new PAGE apparatus


"Dr. James J. Campanella" <[Only registered users see links. ]> schrieb im
Newsbeitrag news:[Only registered users see links. ]. net...

We seal the bottom of the glass plates with parafilm before pouring the gel
(apply some silicone vacuum grease to the edges and then puf the parafilm
on, seals perfectly). it takes about 3 minutes and we didn't have leakage
ever since.



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  #9  
Old 08-10-2005, 04:46 PM
bford
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Default suggestions for new PAGE apparatus

> "Dr. James J. Campanella" <[Only registered users see links. ]> schrieb im

You might consider, given that you are doing large numbers, a gel-block
system such as we use for 2-D gels. Leaks are not an issue.
With a block system, you pour the separation layer for up to 16 gels at
once, inside a closed tank, pumping the gel from the bottom. The gel
plate sets are separated by plastic film so they can be used
individually. Pour the stacking gel after, from the top. We pour about
1.5 litres of gel at once, and the system costs us about 100 ml of that
for hoses and the tank bottom.

You just need to choose a system which uses gels sizes comaptible with
your apparatus, or get one fitted for your system.

Your mileage may vary.

B>




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