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Wanted: Methods for purifying DNA from agarose gels
Andrew Barrett
Friday, 13 April 2007 18:33 UTC
I am relatively new to molecular biology and need to purify DNA from agarose gels. At present, I am phenol:chloroform extracting DNA directly from low melting point agarose – with limited success. I am aware that I could also electro-elute the DNA (followed by phenol:chloroform extraction) or use one of the many gel extraction kits available. I would be grateful for advice on what route to take and what gel extraction kit is the best to use.
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Replies
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Hi Andrew. Do not think too much and go for QIAGEN kit (QIAquick Gel Extraction Kit; 28706). You do not need low-melting point agarose. Just follow the instruction. Good luck! Marek
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I also use the Qiagen gel extraction kits with success.
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The qiagen kit for DNA purification from agarose gels did work well with me.
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I agree with the others…use the kit! Let us know how it worked!
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Our lab also uses the Qiagen gel extraction kit.
Alternatively, you could try cutting bands from regular agarose gels, loading this onto the top of a filter pipette tip with the bottom cut off, load into a 1.5ml tube, spin down for about 10 minutes, remove filter tip, add water to 1.5mil tube, then this should be ready for pcr… obviously this will require some optimization.
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I have used the kit and now have a much better yield of DNA. Many thanks for all your responses, your help is very much appreciated.
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Hi,
You could also try Invitrogen’s gel extrcation kit. Its better than Qiagen.
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Some very useful protocols to be found here:
Electrophoresis onto membranes
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for long DNA fragments, i tend to avoid coljumn kits as the elution is of poor efficiency and rather use matrix purification. I founded also that matrix is less sensitive regarding the amount of agarose used. Th kit i used was Quiagen QiaEX II.
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I have a very simple method to purify DNA from agarose gels. you could cut the specific fragment and put it in a 1.5mL tube, then you chill this tube in liquid nitrogen for about 20s. centrifuge this tube at 13000rpm and get the liquid. add 1/10 volume 3M NaAC and 2-2.5 volume 100* ethanol. next step i think you may be very proficient.
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