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gel analysis of large RNA products


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3 replies to this topic

#1 geneman

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Posted 28 December 2004 - 02:13 PM

I have a large RNA (~6kb) comes from in vitro transcription and labeled with 32P. I runned it by polyacrylmide gel, however, I found that it still stayed in the sample well after I runned at 10mA for 3 h. I also tried to run it on agrose gel and transfer then exposured to x-ray film but without any luck. Did anyone have experience to deal with such large RNA before? What is the best way to visualize it? Thanks.

#2 katika

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Posted 29 December 2004 - 11:50 AM

Did you use formamide-containing agarose gel?

Try etanol precipitation before loading, it is possible that something prevent RNA motility

#3 geneman

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Posted 29 December 2004 - 07:29 PM

Thanks for your kindly reply. I do use the 4% denatured gel. But the bands above 5000 bp of my RNA ladder also stayed in the sample well. Do you think use acrylamide-agrose gel would be helpful? Thanks

#4 katika

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Posted 30 December 2004 - 01:17 PM

Please indicate all conditions of the electrophoresis you perform. May be agarose was too concentrated?




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