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Assay Drug Dev Technol. 2008 Oct;6(5):683-91. doi: 10.1089/adt.2008.142.

Quantitative optimization of reverse transfection conditions for 384-well siRNA library screening.

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  • 1Section of Biologic Research, Department of Biological Technologies, Wyeth Research, Cambridge, MA 02140, USA.


A necessary step in all small interfering RNA (siRNA) library screens is introduction of the siRNA into cells. We describe the use of a commercially available glyceraldehyde 3-phosphate dehydrogenase enzymatic assay that is capable of simultaneously assessing the efficiency of siRNA delivery into cells and the lipid toxicity. This assay has been modified to work in 384-well plates using reverse transfection. The assay is fast, inexpensive, and quantitative. Conditions identified as optimal using this technique have been employed successfully in library screens.

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