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EXtraction of DNA bands from acrylamide gel


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4 replies to this topic

#1 saara

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Posted 01 November 2009 - 02:23 PM

I am trying to extract DNA bands from 4% acrylamide gel, but I had no success. Does anybody thinks that the age of acrylamide gel (my gel stock solution is about a year old) can affect the extraction of DNA bands from the gel.

#2 ShannonJ

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Posted 02 November 2009 - 07:01 AM

In my experience, the acrylamide (if stored at 4C) should be fine. We also never "extracted" DNA from the gels, just cut the bands and placed them into 10mM Tris for 24-48hrs in the fridge. Over that time most of the DNA escaped from the gel into the liquid...at least enough for PCR. Hope that helps.

I am trying to extract DNA bands from 4% acrylamide gel, but I had no success. Does anybody thinks that the age of acrylamide gel (my gel stock solution is about a year old) can affect the extraction of DNA bands from the gel.



#3 Mulletman

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Posted 03 November 2009 - 05:04 AM

In my experience, the acrylamide (if stored at 4C) should be fine. We also never "extracted" DNA from the gels, just cut the bands and placed them into 10mM Tris for 24-48hrs in the fridge. Over that time most of the DNA escaped from the gel into the liquid...at least enough for PCR. Hope that helps.


so we donīt need to use the gel extraction kits?
Awesome cost saver....

#4 ShannonJ

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Posted 03 November 2009 - 07:29 AM

I always have for agarose gels but not for acrylamide...but like I said you had to let 'em sit in the fridge for at least a full day. You might get better recovery with an extraction kit but we were just re-PCRing from DGGE gels and it worked pretty much without fail.

so we donīt need to use the gel extraction kits?
Awesome cost saver....



#5 Qundo12

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Posted 05 November 2009 - 11:08 PM

Did you succeed? I did that in the past. In my case it's the 6% acrylamide. I just cut out the band, place them in water, grind and boil for 15 minutes (the gel won't be dissolved but the acrylamide matrix can be loosen a little bit and DNA can escape to the medium). It's enough for PCR.




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