|Lipo2000 Transfection Reagent Catalog No.GK20005|
Sample solution is provided at 25 µL, 10mM.
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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.
A. Lipo2000 DNA Transfection Reagent Protocol
|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.
|Components||0.75 mL||1.5 mL|
|Lipo2000 Transfection Reagent||0.75 mL||0.75 mL * 2|
|Store at 2–8°C. Do not freeze.|
Lipo2000 Transfection Reagent is a proprietary formulation for transfecting nucleic acids into a wide range of eukaryotic cells. DNA-Lipo 2000 complexes must be made in serum-free medium such as Opti-MEM® Reduced Serum Medium and can be added directly to cells in culture medium, in the presence or absence of serum/antibiotic. It is not necessary to remove complexes or change/add medium after transfection. The amount of Lipo 2000 Reagent required for successful transfection varies depending on the cell type and passage number. Start any new transfection by testing the recommended four concentrations of Lipo2000 Reagent to determine an optimum amount.