Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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RNAi in Drosophila S2 Cells by dsRNA Soaking

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This is a simple method for inducing RNAi in Drosophila S2 cells by soaking the cells in the medium containing dsRNAs. In comparison with transfection (see Protocol 3), soaking requires fewer procedures and avoids the cost of transfection reagents, making it the method of choice for high-throughput RNAi screening. Moreover, soaking avoids potential toxicity associated with transfection reagents. However, delivery of dsRNA to S2 cells by soaking is less efficient than transfection (see Protocol 6), and genes whose expression has proved difficult to repress using soaked dsRNA often can be suppressed by transfecting the dsRNA multiple times. The soaking protocol here is designed for cells grown in six-well plates. If other multiwell plates, flasks, or dishes of a different diameter are used, scale the cell density and reagent volumes according to the surface area of the well (see Table 1).


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(Limited time special offer.) Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook
Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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