How does FuGENE HD work?
How does FuGENE HD work?
FuGENE® HD interacts with nucleic acids to provide efficient and safe entry into the cell, allowing users to overcome barriers in difficult-to-transfect lines such as primary cells, stem cells, and suspension cells. Its simple, easy-to-use protocol allows users to free up valuable lab time.
How do you optimize a siRNA transfection?
To optimize conditions, transfect target cells with several concentrations of an siRNA specific to your chosen positive control and to your experimental target siRNA. Measure the reduction in the control protein or mRNA level compared to untransfected cells 48 hours after transfection.
What is FuGENE transfection reagent?
FuGENE 6 Transfection Reagent is a nonliposomal, multicomponent reagent proven to efficiently transfect more than 700 cell types. It can be used in the presence or absence of serum, and its minimal cytotoxicity eliminates the need to change medium after transfection.
What is the concentration of siRNA for transfection?
1-30 nM
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 the difference between stable and transient transfection?
In stable transfection, the plasmid DNA successfully integrates into the cellular genome and will be passed on to future generations of the cell. However, in transient transfection, the transfected material enters the cell but does not get integrated into the cellular genome.
What is lipofectamine transfection?
Lipofectamine or Lipofectamine 2000 is a common transfection reagent, produced and sold by Invitrogen, used in molecular and cellular biology. It is used to increase the transfection efficiency of RNA (including mRNA and siRNA) or plasmid DNA into in vitro cell cultures by lipofection.
How can I improve my siRNA knockdown efficiency?
- Be Consistent When Conducting Experiments.
- Select Appropriate Order of Transfection.
- Use Healthy Cells at the Optimal Density.
- Choose the appropriate Culture Media and Culturing Conditions.
- Use High Quality siRNA at the Lowest Effective Concentration.
How long is siRNA transfection?
5–7 days
Gene silencing resulting from siRNA can be assessed as early as 24 hours post-transfection. The effect most often will last from 5–7 days. However, the duration and level of knockdown are dependent on the cell type and concentration of siRNA. Transfections may be repeated to maintain silencing.
Is FuGENE toxic to cells?
Fugene was found to be the most toxic reagent for this cell line, with a viability of 68.45%.
What is in FuGENE 6 transfection reagent?
FuGENE® 6 Transfection Reagent is a proprietary blend of lipids and other components supplied in 80% ethanol, sterile-filtered, and pack- aged in glass vials.
What is the purpose of transient transfection?
Transient transfection can be used to investigate the short-term impact of altered gene or protein expression. Transiently transfected genetic material is not integrated into the host genome and therefore the effect on target gene expression is time-limited and is lost during ongoing cell division.
Why choose FuGene® HD transfection?
We offer numerous convenient solutions to meet your lab’s needs. FuGENE® HD Transfection Reagent offers unrivaled transfection performance with minimal impact on cell physiology (Jacobson et al. [2009] Biotechniques 47, 617–24).
Is FuGene® HD a registered trademark?
FuGENE is a registered trademark of Fugent, L.L.C., USA. FuGENE® HD is sold with a Limited Use Label License (LULL). You shall use this Product in accordance with all applicable laws, rules and regulations.
Where can I find transfection conditions for my cell line?
Use the FuGENE® HD protocol database to see if transfection conditions for your cell line are available. The database includes over 25 cancer models and provides transfection protocols specific for each cell line.