DNAfectin Plus Transfection Reagent is a unique formulation of polycations and liposomes, which guarantees high transfection efficiency in a wide variety of cell types including primary cells.
Optimizing Transfection for Specific Cell Lines
To achieve the maximum transfection efficiency and low cytotoxicity, optimize transfection conditions by varying cell density along with DNA and DNAfectin concentrations. Optimal results can be achieved when cells were 70% confluent and DNA (μg): DNAfectin (μl) ratios were 1:3 to 1:5.
| Reagent Quantities for Different Culture Vessels - Specification | ||||
| Vessel | Culture volume | DNA | DNAfectin | Transfection volume |
| 24-well | 500μl | 0.2-0.4μg in 25μl | 0.6-2.0μl in 25μl | 0.3ml |
| 12-well | 1.0ml | 0.5-0.8μg in 50μl | 1.5-4.0μl in 50μl | 0.4ml |
| 6-well | 2.0ml | 1.0-2.0μg in 100μl | 3.0-10μl in 100μl | 1.0ml |
| 35mm | 2.0ml | 1.0-2.0μg in 100μl | 3.0-10μl in 100μl | 1.0ml |
| 60mm | 5.0ml | 3.0-6.0μg in 500μl | 9.0-30μl in 500 | 2.0ml |
| 10cm | 10ml | 8.0-16μg in 1.5ml | 24-80μl in 800μl | 5.0ml |
Note: DNAfectin Plus performs similar to Life Lipofectamine™ 2000 in terms of gene transduction efficiency and cytotoxicity in most cells. In the event that your specific cells are sensitive to this transfection reagent (>30% cells die within 48 hours after transfection), reducing the transfection reagents and the amount of DNA by 50% will solve the problem while achieving successful gene transduction results.
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