siRNA EGFR

Protocol tips

Upstream tips	Protocol tips	Downstream tips
1 × 10^5/well in six-well plates	EGFR siRNA #2 (ID s564), sense 5′-CCAUAAAUGCUACGAAUAUtt-3′ and antisense 5′-AUAUUCGUAGCAUUUAUGGag-3 Add diluted DNA to diluted Lipofectamine® 2000 Reagent and incubate for 5 minutes at room temperature. Incubate cells for 1–3 days at 37°C.

Upstream tips
1 × 10^5/well in six-well plates
Protocol tips
EGFR siRNA #2 (ID s564), sense 5′-CCAUAAAUGCUACGAAUAUtt-3′ and antisense 5′-AUAUUCGUAGCAUUUAUGGag-3 Add diluted DNA to diluted Lipofectamine® 2000 Reagent and incubate for 5 minutes at room temperature. Incubate cells for 1–3 days at 37°C.

Publication protocol

Cells were seeded at a density of 1 × 105/well in six-well plates. On the following day, transfection of small interfering RNA (siRNA) was carried out using Lipofectamine 2000 (Invitrogen). For all procedures, standard protocols were used according to the manufacturers’ manuals.

Synthetic siRNAs targeting EGFR were purchased from Applied Biosystems/Ambion (Foster City, CA). Target sequences were as follows: EGFR siRNA #1 (ID s563), sense 5′-GAAUAGGUAUUGGUGAAUUtt-3′ and antisense 5′-AAUUCACCAAUACCUAUUCcg-3′; EGFR siRNA #2 (ID s564), sense 5′-CCAUAAAUGCUACGAAUAUtt-3′ and antisense 5′-AUAUUCGUAGCAUUUAUGGag-3′; and EGFR siRNA #3 (ID s565), sense 5′-GAUCUUUCCUUCUUAAAGAtt-3′ and antisense 5′-UCUUUAAGAAGGAAAGAUCat-3′. Unspecific siRNA Negative Control #1 (Applied Biosystems/Ambion) was used as control.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for siRNA EGFR below.

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