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Electrophoresis, please help! :(

Electrophoresis, please help! :( - DNA Extraction Forum

Electrophoresis, please help! :( - DNA Isolation Forum. A forum specifically about questions on DNA extraction and purification.


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Old 04-26-2011, 10:06 PM
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Unhappy Electrophoresis, please help! :(



In electrophoresis, between eukaryotic and prokaryotic digests, which would produce the most visible individual bands on agarose gel and why?

I had a practical and this is one of the questions for studying for the following exam. I have realy tried my best to figure it out before I came here

Any help would be much appreciated

Brooke
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Old 04-27-2011, 02:02 AM
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Default Re: Electrophoresis, please help! :(

Prokaryotic should produce a more intense band. Why? Because prokaryotic organisms or cells are smaller than eukaryotic cells, so in the same volume of culture media you could have much more prokaryotic cells than eukaryotic cells, which ultimately result in a bigger yield of DNA per culture and thus a wider band of digest.

However, the above stated do not apply if you digest the same number of cells (i.e. 5 million) in which case the bands should have the same intensity for both kind of cells.
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Old 04-27-2011, 05:10 PM
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Default Re: Electrophoresis, please help! :(

Good point above,
also don't forget there are fewer bands with prokaryotes ie less genes - this will result in fewer but more intense bands
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Old 06-20-2011, 10:46 PM
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Default Re: Electrophoresis, please help! :(

Hi, sorry to bother you all.
I need some help with PCR and DGGE.

I am looking at the soil microbiota as a whole rather than isolating specific bacterial species. In my DGGE profile the intensities of each band is about the same at each lane
(seen at the same row), that's what make my head aches. I was hoping to see a different profile lane to lane, but it seems they were all look the same! But from one band to another in the same lane/column have different intensities. I used 338F-GC and 518R. Any ideas how can I avoid two bacterial species from stopping at the same point in my DGGE gel beside using different primers?
Thanks for your kind answer...

btw, i already make thread at
PCR to DGGE questions
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