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Transfer not occurring on right hand side of membrane correctly


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#1 stelane

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Posted 02 August 2011 - 01:47 PM

Recently we have been experiencing problems with protein transfer using our Invitrogen Surelock system. The right hand side (last four lanes) of the gel are not transferring to the membrane correctly, it is coming out very weak in comparison to the left hand side. If we run the ladder on the right a lot of the dye will remain on the gel, yet will transfer cleanly when loaded on the left. This problem is consistent with all members of the lab. I thought that maybe there could be oxidation on one or both of the metal plates in the transfer box that is impeding current flow at that particular spot, but cannot see any oxidation at all. I have called Invitrogen, but they claim to have not seen this problem before. Has this happened to anyone else, does anyone have any suggestions?

I don't think it is the sandwich construction or sandwich pads as it is always on the right hand side, we always use the same transfer buffer recipe, running times and voltages.

Any help much appreciated.

Edited by stelane, 02 August 2011 - 01:48 PM.


#2 bob1

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Posted 02 August 2011 - 07:18 PM

That's pretty weird. IIRC the surelock system just has a single plate directly connected to the electrode, so there shouldn't be a problem over one side of the tank unless there is a crack in the plate somehow.

Perhaps the tank is leaking slightly or something on that side, so water coming in from outside is diluting the buffer resulting in poorer transfer?

#3 Adrian K

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Posted 03 August 2011 - 08:44 AM

I use the system as well. I have not experience this problem, yet (finger cross).
There was suggestions to submerge the module with nitric acid in the manual to clean it. Try run it again ans see how it goes, and let us know. ^_^

p/s: did you put too many transfer buffer?
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#4 stelane

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Posted 03 August 2011 - 10:14 AM

That's pretty weird. IIRC the surelock system just has a single plate directly connected to the electrode, so there shouldn't be a problem over one side of the tank unless there is a crack in the plate somehow.

Perhaps the tank is leaking slightly or something on that side, so water coming in from outside is diluting the buffer resulting in poorer transfer?


Mate, you are good :rolleyes:!!!!

I just placed the transfer box (empty) into the surelock system, locked it into place and filled the outer chamber with water. As soon as some pressure was placed on the transfer box, a hairline fracture on the bottom right hand side of the transfer box became apparent! Water is slowly leaking into the inner chamber, which I'm guessing would normally run up the transfer pads on the right hand side and interfere with the transfer on that side. Will be trying a new transfer box in the next couple of days, will keep you posted.

Steve




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