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Preparation Before Experiment
1.Plate cells so they will be 70–90% confluent at the time of transfection.
2.Prepare plasmid DNA-lipid complexes.
3.Add DNA-lipid complexes to cells.
Experimental Steps
A. Lipo2000 DNA Transfection Reagent Protocol
| Component | 96-well | 24-well | 6-well |
| Final DNA per well | 100 ng | 500 ng | 2500 ng |
| Final Lipo2000 Reagent per well | 0.2–0.5 μL | 1.0–2.5 μL | 5.0–12.5 μL |
B. Co-Transfection of Plasmid DNA and siRNA
Transfect plasmid DNA and siRNA at the same time using Lipo2000 Reagent by adding 30 pmol (~0.6 μg) of siRNA per 1 μg of DNA.
C. mRNA Transfection
mRNA can be transfected in a 24-well plate using Lipo2000 Reagent by adding 0.5–1 μg of mRNA per well.
D. Lipo2000 DNA Transfection Reagent Protocol
Transfect cells according to the following chart. Volumes are given on a per-well basis. Each reaction mix is sufficient for triplicate (96-well), duplicate (24-well), and single well (6-well) transfections, and accounts for pipetting variations.
