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
