SiRNA Transfection
Reagents:
Lipofectamine 2000 (Invitrogen, Cat# 11668-027) is for plamid DNA transfection.
Lipofectamine RNAiMAX (Invitrogene, Cat# 13778-075 or 13778-150) is proprietary formulation specifically developed for the transfection of siRNA and Stealth RNAi duplexes into eukaryotic cells. High transfection efficiencies, minimal cytotoxicity, requiring low concentration of RNAi duplexes, broad peak of optimal transfection activity.
Oligofectamine (Invitrogen Cat# 12252-011) is proprietary formulation for the transfection of oligonucleotides and short interfering RNA (siRNA) into eukaryotic cells.
Procedures:
1. Plate cells in a 6-well plate at a density of 3x10^5 cells/well.
2. Prepare plasmids： Stock plasmid concentration： 1 ug/ul in TE buffer.
3. Transfect cells at 70-85% confluence with plasmids using Lipofectamine 2000 or Oligofectamine:
2.5 ug plasmid DNA in 150 ul OptiMen, mix gently
10 ul Lipofectamine 2000 in 150 ul OptiMen, mix gently
Mix the above diluted plasmid DNA and diluted Lipofectamine 2000, keep the mixture at RT for 20 minutes
4. Add the plasmid mixture into the cell well. Change media after overnight incubation. Incubate for another 12 hours.
5. Visulize/detect gene expression.
Note: LipoFectamine2000-Instruction
FT201-TransIn-EL-TransfectionReagent-Instruction
