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


A variety of siRNA applications, including siRNA library screening, require the efficient delivery of hundreds to thousands of siRNAs to cells. Most published siRNA studies describe performing transfections in 6 well and 24 well formats. Ambion has developed two reagents, siPORT NeoFX Transfection Agent and siPORT siRNA Electroporation Buffer, and an apparatus, the siPORTer-96 Electroporation Chamber, to facilitate siRNA delivery in 96 well formats. Here we describe experiments analyzing the efficiency and reproducibility of these tools for delivering siRNAs using standard laboratory equipment.

Chemical transfection and electroporation are routinely used to deliver siRNAs into cultured cells. Of these two methods, chemical transfection is most effective for delivering siRNAs into adherent immortalized cell lines, whereas electroporation is recommended for primary cells [16], cells grown in suspension, and other chemical transfection-resistant cell lines. We have adapted both methods for high throughput delivery of siRNAs.


High Throughput Delivery

High throughput siRNA delivery remains a challenge for many laboratories due to the variability of cell culture and delivery efficiency in 96 and 384 well plate formats. There are three important components of a successful siRNA library screen that are directly related to delivery:

Transfection/electroporation effici
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