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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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