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Human rhabdomyosarcoma (RD) cells (CCL136TM, ATCC) cells were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma-Aldrich) enriched with 10% fetal calf serum (FCS, PAA Laboratories) in T75 at 37°C in an atmosphere of 5% CO2. MINK (M-004861–03–0005) siRNA obtained from dharmacon. Specific targeting siRNAs and scrambled siRNAs were dissolved in diethyl pyrocarbonate (DEPC)-treated reverse osmosis (RO) water to a stock concentration of 100μM. The siRNAs were then diluted to desired working concentrations with DharmaFect Cell Culture Reagent (DCCR) and DharmaFect-1 transfection reagent. The siRNAs were transfected into RD cells for 72h at 37°C in a 5% CO2 atmosphere prior to infection. |
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Protocol tips |
Human rhabdomyosarcoma (RD) cells (CCL136TM, ATCC) cells were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma-Aldrich) enriched with 10% fetal calf serum (FCS, PAA Laboratories) in T75 at 37°C in an atmosphere of 5% CO2. MINK (M-004861–03–0005) siRNA obtained from dharmacon. Specific targeting siRNAs and scrambled siRNAs were dissolved in diethyl pyrocarbonate (DEPC)-treated reverse osmosis (RO) water to a stock concentration of 100μM. The siRNAs were then diluted to desired working concentrations with DharmaFect Cell Culture Reagent (DCCR) and DharmaFect-1 transfection reagent. The siRNAs were transfected into RD cells for 72h at 37°C in a 5% CO2 atmosphere prior to infection. |
Publication protocol
Human rhabdomyosarcoma (RD) cells (CCL136TM, ATCC) cells were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma-Aldrich) enriched with 10% fetal calf serum (FCS, PAA Laboratories) in T75 at 37°C in an atmosphere of 5% CO2. MINK (M-004861–03–0005) siRNA obtained from dharmacon. Specific targeting siRNAs and scrambled siRNAs were dissolved in diethyl pyrocarbonate (DEPC)-treated reverse osmosis (RO) water to a stock concentration of 100μM. The siRNAs were then diluted to desired working concentrations with DharmaFect Cell Culture Reagent (DCCR) and DharmaFect-1 transfection reagent. The siRNAs were transfected into RD cells for 72h at 37°C in a 5% CO2 atmosphere prior to infection.
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