Transfectamine™ mRNA Transfection Reagent

1. Prepare cells for transfection.

2. Prepare the Transfectamine™ mRNA Transfection Reagent-RNA mixture.

3. Add the Transfectamine™ mRNA Transfection Reagent-RNA mixture to the cell culture.

4. Culture cells overnight.

5. Analyze transfection efficiency with an appropriate method.

1. Culture cells to ~ 90% confluency at the time of transfection.

2. Replace with fresh growth medium before transfection. For example, replace with 2 mL of medium per well for 6-well plates and 6 mL of medium for 10 cm plates.

1. Mix 2.5 µg of mRNA with 200 µL of serum-free medium.

2. Add 7.5 µL of Transfectamine™ mRNA Transfection Reagent to Step 1.

3. Mix well and incubate at RT for 20 minutes.

   Note: The ratio of Transfectamine™ mRNA Transfection Reagent to mRNA needs to be optimized for different cell lines. In general, Transfectamine™ mRNA Transfection Reagent (µL) to mRNA (µg) Ratio = (3 to 5 µL) to 1 µg.

    

    

Table 1. Sample protocol detail for 6-well plates as shown in the table below.

1. Add the Transfectamine™ mRNA Transfection Reagent -mRNA mixture to the culture plate and culture overnight.

   Note: Recombinant protein expression can be detected as early as 8 hours after the transfection. Maximal expression level may be observed ~24 hours after the transfection.