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Question of transfection reagent

The protocol sent out with Lipofectamine 2000 indicates to use 24 well plates but I am interested in using 60mm plates. How do I scale up my transfection with lipofectamine 2000?

A 60mm plate is 10x greater in surface area than a well from a 24-well plate. Therefore, you would need to scale up all of the components in the reaction by 10x.

The product insert for Lipofectamine 2000 (included with the reagent or can be downloaded from the web site) contains a table indicating the relative surface areas of different cell culture vessels and what factor needed to scale your transfections.

Is Lipofectamine 2000 the same as Lipofectamine?

No. These are two different formulations of lipids and each have been shown to transfect at different efficiencies different cell lines. Please see the Cell Line Database in order to see the optimal usage of either with your cell line.

Is Lipofectamine 2000 stable at room temperature?

Here is information we have regarding stability for this reagent:
LipofectAmine 2000 should be stored at 4oC. It is stable for at least one year.
It is stable for at least 6 months at room temperature if unopened.
It should survive at least one freeze-thaw.
DNA-LF2000 Reagent complexes are stable for at least 6 h at room temperature.

Can Lipofectamine LTX be used in transfection of Suspension cells and other hard to transfect cells?

We have tested transfection of suspension cells such as Jurkat and K562. We achieved transfection efficiency of less than 5% for each of them. We do not notice toxicity with Lipofectamine LTX reagent when used with these suspension cells. For other cell lines please follow the guidelines in the product insert. However if you experience low transfection efficiency, then alternate methods such as electroporation/viral delivery methods are recommended. Please use the following link for cell specific protocols for K562 and Jurkat cells.

How does Lipofectamine LTX compare to Lipofectamine 2000?

Lipofectamine LTX and Lipofectamine 2000 are both recommended for plasmid DNA transfection. However, in a panel of 16 cell lines tested, LTX showed an average of 2.5 fold increase in expression and 25% lower cytotoxicity compared to Lipofectamine 2000.