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High DNA Concentration with Spectrophotometer No Band on Agarose Gel?

High DNA Concentration with Spectrophotometer No Band on Agarose Gel? - Agarose Gel Electrophoresis Forum

High DNA Concentration with Spectrophotometer No Band on Agarose Gel? - Agarose Gel Electrophoresis Forum. A forum to discuss electrophoresis staining, troubleshooting, and development of films for DNA and RNA gels.


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Old 07-05-2008, 04:57 AM
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Question High DNA Concentration with Spectrophotometer No Band on Agarose Gel?



Hi, i got a very good concentration DNA on the UV spectrophotometer but after running the DNA on the gel there was no band on the agarose gel!

Concentration of DNA (420 ng/ul)

Ratio 260/280 nm = 1.17 - 1.47.

Can someone tell me why this happened?

thanks
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Old 07-07-2008, 06:23 PM
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Default Re: High DNA Concentration with Spectrophotometer No Band on Agarose Gel?

With that kind of ratio all you have is protein. Hence no band on gel. proteins absorb at 260 but bind EtBr poorly

Martin
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Old 07-15-2008, 08:17 AM
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Default Re: High DNA Concentration with Spectrophotometer No Band on Agarose Gel?

Probably it may be protein or sometime genome size may be too large, even then that there will be large chunk of single band is possible. What is the percentage of gel? Try lesser percentage of gel? or use some restriction enzymes before going separation.
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Old 07-15-2008, 08:18 AM
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Default Re: High DNA Concentration with Spectrophotometer No Band on Agarose Gel?

Probably it may be protein or sometime genome size may be too large, but even then that there will be large chunk of single band is possible. What is the percentage of gel? Try lesser percentage of gel? or use some restriction enzymes before going separation
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