I have prepared agarose gel, now i want to dissolve the gel completely in order to do the further analysis, I have put in the beaker and added some water to the gel and then I tried stirring at temperature but i couldn't get a homogeneous solution, do some one have a suggestion?
Thank you in advance
In some cases you can use "GELaseTM Agarose", but you will need to use low-melting-point agarose.
https://www.cambio.co.uk/library/images/html_images/Lucigen/gelase-agarose-gel-digesting-preparation-gelase-50x-reaction-buffer.pdf
I suggest this attachment
https://www.goldbio.com/articles/article/AgaroseGelFAQs
I am not really sure why you would attempt to dissolve agarose gel with just water plus heat for the following reasons:
1. By dissolving in just water you will dilute the agarose
2. If you dissolve agarose in water you dilute the ions which are required for electrophoresis
Thus, what you shoukd actually do with agarose that is already in gel form is MELT by placing in a microwave on full power: normally for 2-4 min with periodic stopping and switling of your glass bottle to help with melting
When you made agarose gel initially, am i right in thinking you then disdolved in 1 x TBE or 1 x TAE buffer
Dear NAvidhe
If you have a microwave, you can do it in few minutes!
For details you can see the presentation
ProteoCool n°27 (Overview on Agarose Gel ) on page1\ of my blog:
ProteoCool https://proteocool.blogspot.com/
Ciao
MAnuele
Thank you for your response. Actually I found out that by stirring at the temperature of 100C during about 1h I can dissolve it.
A faster way to quickly have a completely homogenized solution is to make use of a microwave for up to 2 mins (for 1% gel). Agarose gel is normally prepared with TAE or TBE, the TA/TB maintain the pH while the E (i.e EDTA) chelate the ions to prevent DNases activity. I am not sure of the intent behind your choice for water, but you might want to rethink that.
Simply melting in a microwave should solve this problem unless there are other reasons why you would add water in the first instance.
You can simply dissolve agarose gel, first, you have to weigh according to your need and then dissolve in TBE/TAE buffer. Heat it in an Oven for 2min and then you can check if still there is the presence of agarose in particles form then heat it for one more minute till the disappearance of agarose particles and the solution becomes homogenous.
Manuele Martinelli Thanks for your response, it was very useful for me. Also I wonder if you have some methods for characterisation the agarose gel, such as the pore sizes?
Dear Navideh.
I'm sorry but Unfortunately i'm not able to provide you any good advice in this directions. I used a thousand of agarose gel for analize my DNA samples but i never analysed the gel.
good luck
Manuele
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