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Agarose gel to separate btween 4800bp and 4850bp


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3 replies to this topic

#1 alai

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Posted 17 February 2011 - 02:15 AM

Hi there

First post here, so be gentle!

Anyway, i want to separate between the the 2 sizes (4800 and 4850bp) on an agarose gel.

What % gel should be using? I have found conflicting information so i am posting here

Thanks

#2 phage434

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Posted 17 February 2011 - 07:56 AM

I"m sorry to tell you, but this will be almost impossible. Agarose does not have resolution this high. Can you cut the fragment with an enzyme near one end, so that you are looking at the difference between 450 and 500 bases, for example?

#3 Maddie

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Posted 17 February 2011 - 12:35 PM

I"m sorry to tell you, but this will be almost impossible. Agarose does not have resolution this high. Can you cut the fragment with an enzyme near one end, so that you are looking at the difference between 450 and 500 bases, for example?


I agree with phage, I doubt it's possible with agarose.
The use of restriction enzymes is a good idea though B)
Theory is when we know everything and nothing is working. Practice is when everything is working and nobody knows why. Here, we combine theory and practice. Nothing is working and nobody knows why.

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#4 hobglobin

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Posted 17 February 2011 - 01:10 PM

I think Pulsed field gel electrophoresis is the technique you can use, if you have it available.
One must presume that long and short arguments contribute to the same end. - Epicurus
...except casandra's that belong to the funniest, most interesting and imaginative (or over-imaginative?) ones, I suppose.




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