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DGGE - (Jun/09/2001 )

Dear all, I am getting some problems for running a Denaturaning Gradient Gel Electrophoresis. My problem is I can't get a very sharp and distint band on the gel. Is the current and voltage play an important role? I have also tried using a new set of reagents ( acrylamide, temed, APS, TAE buffer, urea...) but its still doesn't work. I will appreaciate for your help. Thank you.




I would like to add a few more questions also. How would the primer dimer produced during PCR affect the DGGE gel run? Is it necessary to purify the PCR product before subjecting to the DGGE gel run (even if the PCR product produce a distinct band without any non-specific product when checking on agarose)?

Many thanks for the help!