Can you use lipofectamine 2000 for siRNA?
Introduction. 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 long is siRNA transfection?
5–7 days
In readily transfected cells treated with potent and effective siRNAs such as the new Ambion Silencer Select siRNAs, near-maximal silencing can be achieved for 5–7 days.
How do you take Lipofectamine 2000?
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.
Does siRNA transfection using Lipofectamine 2000 affect cellular autophagy?
Lipofectamine 2000 is commonly used for siRNA transfections. However, few studies have examined cellular responses to this delivery system. The purpose of this study is to evaluate the effect of siRNA transfection using Lipofectamine 2000 on cellular autophagy. Huh7.5 cells, stably transfected to ex …
How do I use Lipofectamine 2000 to cotransfect my plasmid DNA?
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.
What is the best transfection protocol for Lipofectamine 2000?
Stealth/siRNA Transfection Protocol Lipofectamine 2000 1 Introduction. Highest transfection efficiency in many cell types and formats ( e .g. 2 Protocol – Stealth RNAi or siRNA Transfection. Use this brief procedure to transfect Stealth RNAi or siRNA into mammalian cells in a 24-well format. 3 Scaling Up or Down Transfections.
How do you mix Lipofectamine 2000 with oligomer?
After the 5-minute incubation, combine the diluted oligomer with the diluted Lipofectamine 2000. Mix gently and incubate for 20 minutes at room temperature (solution may appear cloudy). Add the oligomer-Lipofectamine 2000 complexes to each well containing cells and medium.
