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Lipofectamine LTX® Reagent is a proprietary, animal-origin free formulation for the transfection of DNA into eukaryotic cells with low cytotoxicity. This reference provides a recommended procedure to transfect plasmid DNA into HCT-116, human colon carcinoma cells (ATCC no. CCL-247) using Lipofectamine LTX® Reagent.
Important Guidelines for Transfection
Follow these important guidelines when transfecting HCT-116 cells using Lipofectamine LTX® Reagent:
Materials Needed
Have the following reagents on hand before beginning:
Culture vessel | Surface area per well | Volume plating medium | Cells per well | Volume dilution medium | DNA | Lipofectamine® LTX Reagent |
---|---|---|---|---|---|---|
96-well | 0.3 cm2 | 100 μl | 2.5 x 104 | 20 μl | 100 ng | 0.35 - 0.55 μl |
48-well | 1 cm2 | 200 μl | 5 x 104 | 40 μl | 200 ng | 0.7 - 1.1 μl |
24-well | 2 cm2 | 500 μl | 1.25 x 105 | 100 μl | 500 ng | 1.75 - 2.75 μl |
12-well | 4 cm2 | 1 ml | 2.5 x 105 | 200 μl | 1 μg | 3.5 - 5.5 μl |
6-well | 10 cm2 | 2 ml | 6.25 x 105 | 500 μl | 2.5 μg | 8.75 - 13.75 μl |