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help for trouble shooting in 2-D

help for trouble shooting in 2-D - 2-D Gel Electrophoresis Forum

help for trouble shooting in 2-D - 2-D Gel Electrophoresis Forum. A forum dedicated to two dimensional gel electrophoresis, discussion about systems, and troubleshooting its problems.


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Old 04-08-2008, 04:52 PM
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Default help for trouble shooting in 2-D



Hi all,
well i am working on rice proteomics i know most of the people working on proteomics are not from plant scinecs but i think you can help me in trouble shooting.
i run 2-D gel but in 2nd dimension i found that smear is only till upper prtion of gel till about half of the gel.i run same sample in duplicat and situation was same in both gels.there were only 5 or 6 separated spots in center of gel rest of gel was nothing as seems its not even separated.and the point till where gel run was just like line.also i face problem during ruunig gel that mA of ther instrument were decreasing it wasnt stable.but i let the gel to run till bromophenol blue reach till end of plate.what could be the reason if any of you could help me in this.would be great help of you guys.
have good work and best wishes for all of you.
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Old 04-08-2008, 05:32 PM
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Default Re: help for trouble shooting in 2-D

Here's an article that may help, Reduction of proteins during sample preparation and
two-dimensional gel electrophoresis of woody plant
samples ---> [Only registered users see links. ]
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Old 02-15-2009, 05:19 PM
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Default Re: help for trouble shooting in 2-D

hi
Iam working on wheat proteomics. So as u hv said u have problem with resolution of spots, so I think u should use IPG strip pH range 4-7 coz wheat proteins fall in this range. And I think there is no prob with the 2nd dimension as the current falls as the bromophenol reaches towards end coz of depletion of ions. And this is very normal.
Also tell me as to wht are ur focussing conditions.

hope tht I answered ur query.

Regards, naincy
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