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siRNA Transfection of Mouse Bone Marrow-derived Macrophages
Published: Feb 5, 2011 DOI: 10.21769/BioProtoc.28 Views: 24163
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Abstract
Short interfering RNAs (siRNA) are a type of double-stranded RNA molecule, typically 20-25 bp long, that are involved in the phenomenon of RNA interference. siRNA transfection is employed in this protocol to knockdown target gene expression in BMM’phi’ cells. Two days after transfection with cells at 60-80% confluence, the knockdown efficiency can reach 90%.
Keywords: Macrophage
Transfection
SiRNA
Background
Materials and Reagents
- RPMI 1640 medium (RPMI) (Life Technologies, InvitrogenTM, catalog number: 11875-093 )
- Fetal bovine serum (Atlanta Biologicals, catalog number: S10350 )
- Stock penicillin/streptomycin (P/S) (Life Technologies, InvitrogenTM, catalog number: 15140-122 )
- Lipofectamine RNAiMAX (iMAX) (Life Technologies, InvitrogenTM, catalog number: 13778150 )
Equipment
- Cell counter
- 6-well plate
- 24-well plate
Procedure
Category
Immunology > Immune cell function > Macrophage
Molecular Biology > RNA > Transfection
Molecular Biology > RNA > RNA interference
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