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Position: Discussion forums > Bioscience biotechniques discussion forums > Molecular biology troubleshooting forum
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DGGE Troubleshooting
Posted by: LauraZ (IP Hidden, Unregistered user, )
Date: April 12, 2005 08:44AM
Dear all,
I have started 1 month ago to use DGGE for microbial communities analysis in soil and I have some problems with my gels. Actually, I made 2 gels and I had good results (good separation of bands), but I had a problem with the third gel : all the samples that I loaded, stopped between the staking gel and the running gel. It seems that tha DNA can't enter into running gel, but I don't know which could be the problem. Could someone help me, please? Thank you for your help! Bye Laura
Re: DGGE Troubleshooting
Posted by: bassamfahmawi (IP Hidden, Regular member, 42)
Date: April 12, 2005 10:23AM
Hello,
It seems that you have protein contamination in your samples, you might need to try further purify your DNA by removing proteins and try running your gel then. Good Luck!
Re: DGGE Troubleshooting
Posted by: riaz mohammed (IP Hidden, Unregistered user, )
Date: August 17, 2005 05:52AM
Hi,
I have trouble in improving DGGE gel image in terms of sharpness of the bands. My DNA is of good quality and free from protein contamination based on spec readings. PCR products also show up as distinct bands on agarose gels. I am using 30-55% gradient for my samples (rumen samples). My pcr products are of 200 base length. I am preparing my gel with 6% acrylamide and running at 40 V overnight. Could you comment on improving the sharpness of the bands? Thank you
Re: DGGE Troubleshooting
Posted by: Stefan Green (IP Hidden, Unregistered user, )
Date: November 23, 2005 08:39PM
I can refer you to this website for some help. If you have further questions, please send me an email:
[ddgehelp.blogspot.com] Cheers, Stefan
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