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Making a 3% agarose gel.


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#1 spellberg56

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Posted 06 January 2010 - 01:09 PM

Hello. I am preparing a 3% agarose gel and am having difficulty dissolving the agarose. I placed 3 grams of low melt agarose into a flask then added 100 ml of TAE. I then heated it in the microwave and some of the powder stuck to the bottom of the flask and finally after alot of swirlling and heating I got it off. But there was still clumps I couldn't get rid off. Is there an easier way of preparing these high % gels?

Thanks

#2 Gerard

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Posted 06 January 2010 - 01:27 PM

Reverse, first the TAE and then the agarose under swirling and than in the microwave would do the trick.
Ockham's razor
Pluralitas non est ponenda sine necessitate
-- "You must assume no plural without necessity".

#3 klinmed

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Posted 06 January 2010 - 01:33 PM

Hello. I am preparing a 3% agarose gel and am having difficulty dissolving the agarose. I placed 3 grams of low melt agarose into a flask then added 100 ml of TAE. I then heated it in the microwave and some of the powder stuck to the bottom of the flask and finally after alot of swirlling and heating I got it off. But there was still clumps I couldn't get rid off. Is there an easier way of preparing these high % gels?

Thanks


When preparing high concn agarose it is a good idea to 1) slowly add the agarose powder, with mixing, into the buffer (not the other way around) 2) soak the agarose for 15 minutes at room temperature before heating 3) swirl the flask multiple times during the heating to keep the powder suspended.

Alternatively you could use agarose especially developed to resolve small fragments (eg Nusieve 3:1).

Hope this helps.

#4 sanjiun

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Posted 06 January 2010 - 06:34 PM

heat it for awhile in microwave oven, take out, swirl a little bit, heat it again...
and soak agarose in TAE for 15-20 min at room temperature before heating certainly help.

#5 Mazhar Hussain

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Posted 08 January 2010 - 05:38 AM

U can also go as follows:
1) pour agarose in another vessel (Beaker or Flask)
2) Add some TAE into the flask which contain agarose stuck to walls. (TAE should be in measured quantity)
3) Heat it up in Microwave oven.
4) Add agarose in apropriate amount now according to volume of TAE increased.
5) Mix both the solution and microwave them.

Best of luck... :)
Mazhar Hussain
Microbiology & Molecular Genetics
Website: Microbiology On-Line
FaceBook Group: Microbiology On-Line




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