When double-transfecting, do both plasmids always go together? - (Feb/28/2007 )

I've often heard from senior members in my lab that if you transfect to pieces of DNA in parallel, e.g. RNAi + GFP plasmid, the cells that open up always receive both constructs. Then when asked for a reference, the answer is: it's common knowledge. So common that nobody can give a good reference?

How about the experts at BioForum? Have you successfully hunted for a paper on this topic? Would love to read that publication. Let me know.

Best, j

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I havnt found any paper about it but if you transfect 2 plamids with a red and a green fluorescent protien. Check for yourself how many cells have both .

I tried this way and found that 90-95 % cells have both.

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hi, i meet some problem when do co-transfection, the co-transfection efficency is always too lower and only 5-10% cells have both,i using lipofectamin2000 and both plasmids concentration is good, I have tried much times but never get good result,it's so frustrated. could you give me some detail information to improve the co-transfection efficiency? thanks ,my email is bobxiang4@hotmail.com, any suggestion is appreciated.

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Thanks for the answer scolix. Did you electroporate? See bobxiang4's comment. Maybe co-transfection has different outcome with amphiphilic transfection reagents? Or did you try those as well in the red/green test?

~j

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I have transfected using lipofectamine and calcium phosphate with red and green fluorescent protiens.

Also if you have viruses expressing either red or green fluorescent protien, inject them together, you will again find a similar result. both invitro and invivo.

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