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EMSA-Difficulties - no clean lanes (Jun/26/2007 )

Hi All,

I have been running a 5% gel for my EMSAs and have been seeing some smear on my gel. We don`t see this smear in the lanes with only probes. When I took some control samples of somebody else, we didn`t saw the smear there. So I think it has to be something with my nuclear extract?! mellow.gif The smear is most of the time located around the shift. (see attachment)[attachment=3198:EMSA__25juni07.ppt]
Can someone tell me what the problem is? huh.gif Or does someone have some experience with it? sad.gif

Please let me know what you all experts think about this problem! blush.gif

Thanks in advance,



I could be wrong but it may be the gel dryer. Is it working efficiently and is it cold before you start ?
If you think it's the extract maybe try titrating the extract down.

All the best,