Can you use Lipofectamine 2000 for siRNA?
invitrogen Lipofectamine 2000 Reagent is a proprietary formulation that facilitates highly efficient delivery of Invitrogen Stealth RNA molecules, short interfering RNA (siRNA) or plasmid DNA to mammalian cells for RNAi analysis (1, 2).
How much siRNA do you use for transfection?
As mentioned above, too much siRNA may lead to off-target effects; too little can result in undetectable gene silencing. In general, 1-30 nM siRNA is a good concentration range within which to optimize transfection (10 nM is a sufficient starting point).
What is transfection with siRNA?
Small interfering RNA (siRNA), also known as silencing RNA, is a double-stranded segment of RNA that can perform various functions in a biological system. A siRNA transfection is the insertion of siRNA into a cell, a process that can be invaluable to gene silencing experiments.
How do you take Lipofectamine 2000?
Nucleic acid-Lipofectamine 2000 complexes can be added directly to cells in culture medium, in the presence or absence of serum. It is not necessary to remove complexes or change/add medium after transfection, but complexes may be removed after 4-6 hours.
What concentration should I use for siRNA?
In general, 1-30 nM siRNA is a good concentration range within which to optimize transfection (10 nM is a sufficient starting point). In Figure 6, transfection of HeLa cells was optimized at very low concentrations of siRNA.
What is the difference between Lipofectamine 2000 and 3000?
Lipofectamine 3000 reagent yields higher transfection efficiencies than Lipofectamine 2000 reagent when tested in a variety of cell lines.
What is siRNA control?
Our siRNA controls allow you to: Determine the role of non-specific cellular responses in your phenotype. Achieve greater knockdown by optimizing transfection conditions. Ensure ongoing experimental success.
How does siRNA drug work?
siRNAs can alter or stop the production of disease-causing proteins by harnessing RNA interference (RNAi), a process that occurs naturally within our cells. RNAi was first discovered by Craig Cameron Mello and Andrew Fire, who won the 2006 Nobel prize for Physiology and Medicine for their groundbreaking work.
When to use Lipofectamine 2000 in cell transfection?
Follow these general guidelines when using Lipofectamine 2000 to cotransfect your plasmid DNA and the RNAi molecule of interest into mammalian cells. Use low-passage cells, and make sure that cells are healthy and greater than 90% viable before transfection. Transfect cells at 80-90% confluence.
How to make Lipofectamine 2000 with siRNA oligomer?
A. Dilute 20 pmol Stealth RNAi or siRNA oligomer in 50 µl Gibco™ Opti-MEM™ I Reduced Serum Medium without serum (resulting concentration of RNA in Opti-MEM is 400 nM). Mix gently B. Mix Lipofectamine 2000 gently before use, then dilute 1 µl in 50 µl OptiMEM I Reduced Serum Medium. Mix gently and incubate for 5 minutes at room temperature.
How to use Lipofectamine 2000 for gene knockdown?
Add the oligomer-Lipofectamine 2000 complexes to each well containing cells and medium. Mix gently by rocking the plate back and forth. Incubate the cells at 37°C in a CO2 incubator for 24-96 hours until you are ready to assay for gene knockdown. Medium may be changed after 4-6 hours.
How to mix Lipofectamine 2000 with Opti-Mem I medium?
Mix gently. Mix Lipofectamine 2000 gently before use, then dilute the appropriate amount in Opti-MEM I Medium without serum. Mix gently and incubate for 5 minutes at room temperature. After the 5 minute incubation, combine the diluted DNA and RNAi molecule with the diluted Lipofectamine 2000.