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TransMessenger Transfection Reagent Handbook

s 2 g. A pipetting scheme for optimizing the transfection of adherent cells in 6-well plates is provided in Table 2, page 11. For transfection using other culture formats, please refer to Table 3

Amount of Enhancer R
The RNA (g) to Enhancer R (l) ratio of 1:2 given in the pipetting scheme on page 11 should not be changed. Efficient condensation of RNA with Enhancer R is determined by the mass quantity of RNA.

Ratio of TransMessenger Transfection Reagent to RNAEnhancer R mixture
The overall charge of the TransMessengerRNA complex is determined by the ratio of TransMessenger Transfection Reagent to RNAEnhancer R mixture. Optimal binding of TransMessengerRNA complexes to negatively charged groups (e.g., sialylated glycoproteins) on the cell surface requires a slightly positive net charge. The ratio of TransMessenger Transfection Reagent (l) to RNA (g) is an important factor to optimize for every new cell type and RNA used. As a starting point for optimization we recommend using an RNA:TransMessenger Transfection Reagent ratio of 2 g RNA to 8 l TransMessenger Reagent when using 6-well plates. A pipetting scheme for optimizing the transfection of adherent cells in 6-well plates is provided in Table 2. To optimize transfection in other culture formats, prepare separate transfection mixtures using:
The starting point RNA and TransMessenger Transfection Reagent quantities listed in Table 3,
Roughly half the listed ratios and quantities
Double the listed ratios and quantities.

Incubation period with TransMessengerRNA complexes
Remove TransMessengerRNA complexes 3 hours after their
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