Table of Contents
What is transfection with siRNA?
Summary of siRNA Transfection 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.
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).
What is the function of siRNA?
siRNAs. siRNAs are highly specific and usually synthesized to reduce the translation of specific messenger RNAs (mRNAs). This is done to reduce the synthesis of particular proteins. They form from double-stranded RNA transcribed and then cut to size in the nucleus before releasing into the cytoplasm.
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 lipofectamine facilitates the entry of DNA into cells?
Lipid-mediated transfection relies on cationic lipids, such as lipofectamine, which facilitate DNA and mRNA delivery into cells. The cationic lipids form a unilamellar liposomal structure with a positive surface charge and fuse with nucleic acids to form a transfection complex.
What is siRNA technology?
The underlying molecular mechanism of gene silencing provides us with short interfering RNAs (siRNAs) which allows to target any gene with high specificity and efficiency. siRNAs can now be obtained in various ways allowing for numerous in vitro and in vivo applications.
How do you use siRNA?
Top 4 ways to make your siRNA experiment a success
- Choose appropriate siRNA reagents and delivery methods that are adapted to your cell type.
- Use appropriate positive and negative controls in each and every experiment.
- Optimize siRNA delivery conditions.
- Confirm your knockdown on the mRNA level using qPCR.
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 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 Invitrogen Lipofectamine 2000 reagent?
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 do you transfect plasmid DNA with siRNA?
Co-Transfection of Plasmid DNA and siRNA Transfect plasmid DNA and siRNA at the same time using Lipofectamine®2000 Reagent by adding 30 pmol (~0.6 μg) of siRNA per 1 μg of DNA. mRNA Transfection mRNA can be transfected in a 24-well plate using Lipofectamine®2000 Reagent by adding 0.5–1 μg of mRNA per well. Photograph of Expected Results