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Transfection Agent


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#1 altobarn

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Posted 08 August 2009 - 09:46 AM

I have previously used Lipofectamine 2000 for my luciferase plasmid transfection and it works perfectly well with merely 1ul in each 24well of 500ul. However in my last transfection I tried Lipofectamine (not with the '2000' version) and when I measure the luciferase, I do not get good readings but just background readings. Im now suspecting the without 2000 Lipofectamine version was the culprit,as I did not change any protocol or chemicals except the transfection agent.

Anyone out there got a clue what went wrong with my luciferase measurement? What are the difference between Lipofectamine 2000 and Lipofectamine without 2000?

Thanks in advance!

#2 Dr Teeth

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Posted 08 August 2009 - 11:19 AM

I have previously used Lipofectamine 2000 for my luciferase plasmid transfection and it works perfectly well with merely 1ul in each 24well of 500ul. However in my last transfection I tried Lipofectamine (not with the '2000' version) and when I measure the luciferase, I do not get good readings but just background readings. Im now suspecting the without 2000 Lipofectamine version was the culprit,as I did not change any protocol or chemicals except the transfection agent.

Anyone out there got a clue what went wrong with my luciferase measurement? What are the difference between Lipofectamine 2000 and Lipofectamine without 2000?

Thanks in advance!


There is a big difference between lipofectamine and lipofectamine 2000. They tend to be used to transfect different cell lines (though not many people use regular lipofectamine anymore) with lipofectamine 2000 being a bit broader in range of cell lines and better in terms of transfecting hard-to-transfect cell types. Each reagent would need to be optimized before use (i.e. the protocol (lipo/DNA ratio) that works best for one reagent in one cell line may not work best with the other reagent or in a different cell line) and the same absolutely will NOT work if you are transfecting in the presence of serum. While lipofectamine 2000 can be used in the presence or absence of serum, lipofectamine must be used in the absence of serum.

Edited by Dr Teeth, 08 August 2009 - 11:21 AM.


Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
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