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Huge Smear in EMSA

emsa

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#1 ugasweet

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Posted 25 November 2013 - 12:20 PM

I am using biotin labeled oligos and I am getting just a giant smear all the way down the lane. I have tried diluting the amount of biotin labeled oligo I'm using and while the intensity of the smear decreases, there is still a smear down the entire lane. I can tell that there are bands at certain places but I would like to clean it up. I am using 2ug protein and poly di/dc. Would it be helpful to add in other components to the binding reaction or should I continue to try diluting my biotin labeled oligos?

 

Any help is greatly appreciated!!



#2 pcrman

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Posted 10 December 2013 - 10:17 PM

How about the lane with oligo minus proteins - do you get similar smear? Are you using nuclear extracts or other protein? 







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