poor polymerization in DGGE

Rafael Maldonado rafamaldo at yahoo.com
Mon May 9 04:41:13 EST 2005


WU Yucheng wrote:
> Hi everyone,
> 
> I am investigating the bacteria diversity in soil by DGGE method.But a
> poor polymerization always happened during casting. In fact,the
> acrylamide solution doesn't polymerize from below the comb about 1cm to
> the top even overnight at temperature(about 20¡æ). The gel oncentration
> is 8% and 10ul TEMED and 150ul 10%APS are added to 15ml high or low gel
> solution.
> 
> Then,who can tell me what is the reason and how I can optimize the
> polymerization.
> 
> Thank you in advance for your help!!
> 
> yucheng wu
> 
> _________________________________________________________________
> Ãâ·ÑÏÂÔØ MSN Explorer:   http://explorer.msn.com/lccn/ 
> ---

Most of the time, poor polymerization is due to oxygen (from the air, if
the buffer is shaked) or APS/TEMED in bad condition. Use new bottles of
APS/TEMED. De-gasing your gel prior the addition of APS/TEMED could be
useful: use a suction flask with a rubber cap and apply vacuum for half
an hour.

-R



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