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DNA extraction


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#1 toniX

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Posted 01 July 2010 - 04:50 PM

hi mates,
I have to separate and extract DNA from a gel. The problem comes because the insert is 4.5kb and the plasmid 4kb. Do I have to use a .7% or 1% agarose? is it alright to use TAE or is better for this case the TBE?
thanks in advance
-t

#2 phage434

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Posted 01 July 2010 - 05:54 PM

Your best bet would be to find an enzyme that cuts the vector but not the desired insert. Then you would have an easy time with the gel. I would use TAE and a 1% gel.
You might want to run it slowly to reduce heat and diffusion.

#3 toniX

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Posted 01 July 2010 - 06:33 PM

Your best bet would be to find an enzyme that cuts the vector but not the desired insert. Then you would have an easy time with the gel. I would use TAE and a 1% gel.
You might want to run it slowly to reduce heat and diffusion.

thanks
I already did a test and the problem was the separation of the two bands (I have to admit that I used a .7% gel and I ran it like there will not be tomorrow, at 150V). I will follow your indications
-t




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