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High Throughput siRNA Delivery In Vitro: From Cell Lines to Primary Cells

ency maximizes reduction in target gene expression and improves the likelihood that a significant phenotype will be observed.

Reproducibility of siRNA delivery between wells and between plates reduces the variability of quantitative phenotypic results and improves the resolution of screening data.

Cell viability increases specificity of the siRNA effect, further improving the likelihood that significant phenotypic differences will be observed.


High Throughput Chemical Reverse Transfection--For Immortalized Cells

We adapted reverse transfection (a variation of standard transfection that involves transfection in suspension; see "Reverse Transfection" at right) to facilitate high throughput siRNA delivery. Because the procedure features minimal pipetting steps and can be performed using multi-channel pipettors, reverse transfection can be used to deliver siRNAs to approximately 1000 cell samples per hour without robotics. It is also amenable to automation

Efficiency of High Throughput Reverse Transfection. To assess transfection efficiency, we mixed different concentrations of a well-characterized GAPDH siRNA with the versatile siPORT NeoFX Transfection Agent in wells of a 96 well plate. Five different cell types were added to the siRNA/lipid complexes to effect reverse transfection.
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