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Electrophoretic Mobility Shift Assay Forum EMSA Forum. Discuss the Electrophoretic Mobility Shift Assay protocols or technique and the assessment of DNA and RNA binding proteins here. Also post questions and troubleshooting about the shift assay.


EMSA with PCR fragments

EMSA with PCR fragments - Electrophoretic Mobility Shift Assay Forum

EMSA with PCR fragments - EMSA Forum. Discuss the Electrophoretic Mobility Shift Assay protocols or technique and the assessment of DNA and RNA binding proteins here. Also post questions and troubleshooting about the shift assay.


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Old 05-23-2008, 04:46 PM
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Question EMSA with PCR fragments



Hi,

Has anyone tried doing EMSA with PCR fragments of a promoter, instead of oligos?

Thanks!
T84
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Old 09-04-2008, 10:29 PM
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Default Re: EMSA with PCR fragments

Hi, yes this is possible however you must label your PCR products after PCR. This is not easily done however if you digest or somehow alter the DNA to have ends restriction digested you can easily Klenow in p32-Atp or other nucleotide in. Also, its been known to be done with biotin.


There was a method to biotinylate DNA fragments even undigested after PCR but I forget now its been a while... I think it used a special enzyme.
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