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The amount of plasmid for ligation reaction


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#1 widget

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Posted 17 February 2005 - 08:21 AM

First time poster here. I have done a transfection today and I wanted to know if the results will be greatly affected by the addition of less or more plasmid DNA than the protocol recommends? Does anyone have any experience with this?

#2 Simonsays

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Posted 17 February 2005 - 01:08 PM

You always have to adapt all conditions depending on DNA lenght, type of transfectant, cell type, #of cells...
I suggest you do some controls by transfecting GFP and try different conditions (g of plasmid, l of transfectant).

Simon

#3 fred_33

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Posted 19 February 2005 - 03:06 AM

hi
i'm still working with the very old lipofectamine reagent (yes it still exists :)) and on the reference guide they give a graphical analysis on the efficiency of transfection. You can go to the page at :
http://molecool.wust...-Invitrogen.pdf
(it's not the one from invitrogen because they've upgraded the notice and didn't keep the table).
But Simonsays is right. You have to adapt conditions for your experiments...




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