X-tremeGENE™ transfection reagents provide reliable transfection of diverse nucleic acids and CRISPR/Cas9 components, a critical step for your molecular and cellular research. Explore our list of the most common cell lines below for detailed instructions on using the X-tremeGENE™ portfolio. Looking for maximal flexibility? The new X-tremeGENE™ 360 transfection reagent is a universal polymer that delivers excellent performance for both commonly used and hard-to-transfect cell lines.
Plate CHO-K1 cells at a density of 1.5 X 104 cells/well. Plate cells in a volume of 100 μL complete growth medium per well in a 96-well plate 18 – 24 hours before transfection (65 – 75% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ 9 DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Experimental result: CHO-K1 cells were successfully transfected with pcDNA3.1-GFP plasmid using using X-tremeGENE™ 9 DNA Transfection Reagent.
Plate COS-7 cells at a density of 8.0 – 9.0 X 103 cells/well. Plate cells in a volume of 100 μL complete growth medium per well in a 96-well plate 18 – 24 hours before transfection (75 – 85% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ 9 DNA Transfection Reagent, DNA and diluent (Opti-MEMR® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Experimental result: COS-7 cells were successfully transfected with pcDNA3.1-GFP plasmid using X-tremeGENE™ 9 DNA Transfection Reagent.
Plate HeLa cells at a density of 1.5 X 104 cells/well. Plate cells in a volume of 100 μL complete growth medium per well in a 96-well plate 18 – 24 hours before transfection (70 – 90% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ 9 Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Experimental result: HeLa cells were successfully transfected with pcDNA3.1-GFP plasmid using X-tremeGENE™ 9 DNA Transfection Reagent.
Plate NIH-3T3 cells at a density of 1.5 X 104 cells/well. Plate cells in a volume of 100 μL complete growth medium per well in a 96-well plate 18 – 24 hours before transfection (70 – 80% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ 9 DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Experimental result: NIH-3T3 cells were successfully transfected with pcDNA3.1-GFP plasmid using X-tremeGENE™ 9 DNA Transfection Reagent.
Plate HEK-293 cells at a density of 3.0 – 3.5 X 104 cells/well. Plate cells in a volume of 100 μL complete growth medium per well in a 96-well plate 18 – 24 hours before transfection (50 – 70% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ HP DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Experimental result: HEK-293 cells were successfully transfected with pcDNA3.1-GFP plasmid using X-tremeGENE™ HP DNA Transfection Reagent.
Plate PC-3 cells at a density of 1.0 – 1.2 X 104 cells/well. Plate cells in a volume of 100 μL complete growth medium per well in a 96-well plate 18 – 24 hours before transfection (75 – 85% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ HP DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Add 2 μL X-tremeGENE™ HP DNA Transfection Reagent to the diluted DNA. (1:1 ratio of reagent to DNA). Pipet gently to mix.
Incubate for 15 – 30 min at +15 °C to +25 °C.
Experimental result: PC-3 cells were successfully transfected with pcDNA3.1-GFP plasmid using X-tremeGENE™ HP DNA Transfection Reagent.
Plate HCT 116 cells at a density of 3.0 – 4.0 X 105 cells/well. Plate cells in a volume of 1 mL complete growth medium per well in a 12-well plate 18 – 24 hours before transfection (80% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ HP DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Plate A549 cells at a density of 2.8 X 104 cells/well. Plate cells in a volume of 150 μL complete growth medium per well in a 96-well plate 18 – 24 hours before transfection (70% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ HP DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Place 500 μL diluent in a sterile tube.
Add 5 μg plasmid DNA. Pipet gently to mix.
Plate MCF-7 cells at a density of 1.2 – 1.8 X 104 cells/well. Plate cells in a volume of 100 μL complete growth medium per well in a 96-well plate 18 – 24 hours before transfection (70 – 90% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ 9 DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Experimental result: MCF-7 cells were successfully transfected with pcDNA3.1-GFP plasmid using X-tremeGENE™ 9 DNA Transfection Reagent.
Plate HuH-7 cells at a density of 1.0 X 105 cells/well. Plate cells in a volume of 2.5 mL complete growth medium per well in a 6-well plate 18 – 24 hours before transfection. Incubate cell cultures overnight.
Allow X-tremeGENE™ HP DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Plate U-2 OS cells at a density of 2.5 – 3.0 X 104 cells/well. Plate cells in a volume of 100 μL complete growth medium per well in a 96-well plate 18 – 24 hours before transfection (70 – 90% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ HP DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Experimental result: U-2 OS cells were successfully transfected with pcDNA3.1-GFP plasmid using X-tremeGENE™ HP DNA Transfection Reagent.
Plate HepG2 cells at a density of 2.0 – 2.5 X 104 cells/well. Plate cells in a volume of 100 μL complete growth medium per well in a 96-well plate 18 – 24 hours before transfection (60 – 70% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ 9 DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Experimental result: HepG2 cells were successfully transfected with pcDNA3.1-GFP plasmid using X-tremeGENE™ 9 DNA Transfection Reagent.
Protocol provided by a customer.
Plate Neuro-2a cells at a density of 1.0 X 105 cells/well. Plate cells in a volume of 500 μL complete growth medium per well in a 24-well plate 24 hours before transfection (30 – 40% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ 9 DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Note: Data and experimental conditions included in “Protocols provided by customers” are the sole responsibility of the customers that have submitted them. Roche was neither involved in establishing the experimental conditions nor in defi ning the criteria for the performance of the specifi c assays. Roche therefore cannot take any responsibility for performance or interpretation of results obtained for the biological target parameter(s) described by the authors or other users using a similar experimental approach.expression.
Experimental result: Neuro-2a cells were successfully transfected with CMV6-GFP plasmid using X-tremeGENE™ 9 DNA Transfection Reagent.
Plate primary MEF cells at a density of 0.5 – 1.0 X 105 cells/well. Plate cells in a volume of 1 mL complete growth medium per well in a 12-well plate 18 – 24 hours before transfection (60% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ HP DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Plate human primary fibroblast cells at a density of 1.0 X 105 cells/well. Plate cells in a volume of 2 mL complete growth medium per well in a 6-well plate 18 – 24 hours before transfection (40 – 50% confl uency). Incubate cell cultures overnight.
Allow X-tremeGENE™ HP DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Experimental result: Human primary fibroblasts were successfully transfected with GFP-encoding plasmid using X-tremeGENE™ HP DNA Transfection Reagent.
Plate EK8 cells at a density of 3.0 X 104 cells/well. Plate cells (3 X 105 cells/mL) in a volume of 100 μL complete growth medium per well in a 96-well plate 24 hours before transfection (50% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ HP DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Plate hMSCs at a density of 8.0 X 104 cells/well. Plate cells in a volume of 2 mL complete growth medium per well in a 6-well plate 18 – 24 hours before transfection (50% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ HP DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Experimental result: human mesenchymal stem cells were successfully transfected with GFP-encoding plasmid using X-tremeGENE™ HP DNA Transfection Reagent.
Plate HEK-293T cells at a density of 5.0 X 105 cells/well. Plate cells in a volume of 2 mL complete growth medium per well in a 6-well plate 18 – 24 hours before transfection (60 – 80% confluency). Incubate cell cultures overnight.
Allow X-tremeGENE™ HP DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Experimental result: HEK-293T cells were successfully transfected with pGIPZ-eGFP plasmid using X-tremeGENE™ HP DNA Transfection Reagent.
Plate SF9 cells at a density of 0.5 X 106 cells/well. Plate cells in a volume of 1 mL complete growth medium per well in a 12-well plate immediately before transfection. Incubate cell cultures overnight.
Allow X-tremeGENE™ HP DNA Transfection Reagent, DNA and diluent (Opti-MEM® I Reduced Serum Medium or serum-free medium) to warm to +15 °C to +25 °C, and vortex gently.
Please be aware that the protocols are suggestions and that optimal conditions must be determined empirically.
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