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DGGE problem - (Apr/28/2005 )

Hi, everyone.

I am doing some denature gradient gel electrophesis. I have a problem with one PCR product- about 360bp. Some of the DNA remain in the stacking gel. But I tried to run smaller PCR product- about 120bp, there is no such problem. I add 40ul 10%APS and 20ul TEMED into 10ml 0% denaturant (8% acrylamiade). Then I tried to use less APS and TEMED, but it seems no difference to 360 bp PCR product. Any idea how can i get rid of the DNA in the stacking gel? How much APS and TEMED you are using noemally?

many thanks for any information.


Afraid my days of DGGE are far in the past (thank god! tongue.gif ), but try posting to this group:

You have to register first. They're usually very helpful.
Good luck!