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RNA contamination in DNA samples


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#1 Lfs

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Posted 08 August 2013 - 07:24 AM

Hi!

 

I would like to know if a DNA sample with RNA contamination can be used for transfecting B16, 293H and SH SY5Y cells.

 

 

Thanks for the help!



#2 pcrman

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Posted 14 August 2013 - 12:31 PM

Is the DNA sample plasmid? I don't think that will be a problem. 



#3 Lfs

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Posted 14 August 2013 - 03:11 PM

Yes, it is!

 

I thought that only protein/lipids contamination could interfere in this method, but my transfection (using the RNA contaminated DNA sample) didn't work unsure.png. In parallel, I transfected B16 cells with another plasmid with a EGFP tag (without contamination) and it worked well. And to make things worse, when I used both plasmids, neither the EGFP nor the RNA contaminated worked. blink.png

 

Maybe I could try a RNAse treatment (again) following by phenol/chlorof extration and ethanol precipitation in order to purify my DNA (because I usually do home made Midiprep dry.png Oh God, I need thoseMidi/Maxiprep Kits!!)...  What do you think about it?

 

Thank you!






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