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how to separate DNA of similar size


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#1 wcnb

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Posted 01 November 2004 - 01:43 PM

Hello, everyone:

I've got 6 DNA fragments range from 600-750 bp and would like to get a optimum condition to separate them best. I tried 1.0%-2.0% agarose gel in TAE buffer with 80V but the diference is not clear enough. Would you please give me some suggestion? Thank you in advance.

#2 rassen

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Posted 01 November 2004 - 03:59 PM

I think 2% gel should work or you can also try polyacrylamide gel.

#3 bob1

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Posted 01 November 2004 - 07:34 PM

You could try a density gradient separation (e.g. using cesium chloride), but I think that your products may be a little small for this, you would have to calculate the density required to separate them and thus the amount of cesium chloride

#4 chandima

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Posted 02 November 2004 - 03:37 AM

hi
i think >1.5% should work it has worked for me.

#5 wcnb

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Posted 02 November 2004 - 06:16 AM

chandima, on Nov 2 2004, 04:37 AM, said:

hi
i think >1.5% should work it has worked for me.
Hi, Chandima,

Thanks for your and others' replies.

would you please tell me the detailed condition you used, like the concentration of gel, the amount of voltage, the type of agarose gel, the buffer. Thanks.

#6 Charon

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Posted 02 November 2004 - 06:30 AM

1-,18% (v/v) Agarose in either TB or TA buffer (TB has better capacity If iI remember corectly)
50-80 V




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