Recent replies to "Wanted: Methods for purifying DNA from agarose gels"

Reply from Mingyong Xiong

Mingyong Xiong — Wed, 05 Sep 2007 04:01:05 -0000

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.

Reply from Frederic Guilhem

Frederic Guilhem — Tue, 10 Jul 2007 11:30:57 -0000

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.

Reply from Chunk Style

Chunk Style — Mon, 18 Jun 2007 20:43:44 -0000

Some very useful protocols to be found here:

Organic Extraction

Electrophoresis onto membranes

Electroelution into dialysis bags

Digestion with Agarase

Reply from Malik John Mohammed

Malik John Mohammed — Fri, 04 May 2007 14:52:33 -0000

Hi,

You could also try Invitrogen’s gel extrcation kit. Its better than Qiagen.

Reply from Andrew Barrett

Andrew Barrett — Tue, 24 Apr 2007 19:43:16 -0000

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.

Reply from Teresita McLenon-Porter

Teresita McLenon-Porter — Tue, 24 Apr 2007 15:31:25 -0000

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.

Reply from Barbara Xella-Pontecorvi

Barbara Xella-Pontecorvi — Wed, 18 Apr 2007 14:02:52 -0000

I agree with the others…use the kit! Let us know how it worked!

Reply from smitha karunakaran

smitha karunakaran — Wed, 18 Apr 2007 00:47:50 -0000

The qiagen kit for DNA purification from agarose gels did work well with me.

Reply from Paul Cullen

Paul Cullen — Tue, 17 Apr 2007 04:52:05 -0000

I also use the Qiagen gel extraction kits with success.

Reply from Marek Cebecauer

Marek Cebecauer — Sat, 14 Apr 2007 15:08:40 -0000

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