Dispersion of Agarose

afc at gnv.ifas.ufl.edu afc at gnv.ifas.ufl.edu
Mon Jan 11 09:15:33 EST 1993


In article <01GTD30FXAKG0000FE at irri.cgnet.com>, RSCOTT at IRRI.CGNET.COM writes:
> I can't help cursing myself whenever I prepare agarose gels and
> the suspension happens to overboil inside the microwave. At
> certain equally irksome times (especially with bigger preparations)
> the gel remained undispersed while considerable volume has already
> been lost during heating. I always cover the boiling flask
> with a beaker but this didn't do so much to prevent excessive 
> evaporation. I discovered a trick (perhaps I just rediscovered it on
> my own) to avoid these pitfalls. I tried presoaking the gel overnight 
> at the eve of my gel run. Wholla! The agarose dispersed quicker in the 
> buffer. Dispersion took only about half the normal boiling time. By the 
> time vigorous boiling commences the agarose particles were substantially
> dissolved. Volume loss is therefore minimized. Presoaking allows 
> the gel to swell and be hydrated. Alternatively, one can equilibrate
> the gel suspension in a 55-650C bath (ca. 30 min) prior to boiling.
> The same result is achieved. I think it's not much trouble having to
> prepare the suspension well in advance and stand it for a while
> especially if the gel consistency can be maintained for different
> preparations of equal concentration.
> 
> R. P. Scott
> Pathology Division
> IRRI
> Manila, Philippines
> E-Mail: rscott at irri.cgnet.com

One caveat:  If you are using a buffer such as tris acetate, this might be 
an excellent way to grow microorganisms.  That could lead to all sorts of
unpleasant and hard to replicate results.

A good suggestion if you are running tris borate gels.

Andrew Cockburn
USDA



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