Table of Contents
How does FuGENE 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.
What is FuGENE transfection reagents and how does it work?
FuGENE® HD Transfection Reagent is a multi-component reagent that forms a complex with DNA, then transports the complex into animal or insect cells. It is suitable for use in media with or without serum, and for transient or stable transfection, as well as co-transfections of multiple DNA plasmids.
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 PEI transfection?
As with most transfection reagents, PEI-mediated transfection involves the formation of nanoparticles (polyplexes) which are obtained by its mixing with plasmid DNA. A short incubation period that allows polyplexes to reach their optimal size is performed prior to their addition to the culture.
Why is transient transfection important?
Transfection, the process of introducing foreign genetic material into a eukaryotic cell, is an important tool for many cell and molecular biologists, as well as anyone studying the effects of altered gene expression on cellular physiology.
How do you make a PEI solution?
Preparation of PEI Stocks
- Dissolve 100 mg in 100 mL sterile ddH2O.
- Stir while slowly adding HCl to pH 7.0.
- Mix for 10 minutes and then recheck pH.
- Filter sterilize through 0.22 um filter.
- Aliquot 500 uL to 1000 uL and store in -80C.
- Thawed solution can be stored at 4C for up to 2 months, label a tube when thawed.
Is transduction better than transfection?
The main difference between transfection and transduction is that transfection is the transfer of DNA without using a virus as a vector whereas transduction is the transfer of DNA with the use of a viral vector. Transfection uses chemical and non-chemical based methods to transfer foreign DNA into the cells.
Why do we use transfection?
The main purpose of transfection is to study the function of genes or gene products, by enhancing or inhibiting specific gene expression in cells, and to produce recombinant proteins in mammalian cells [3].
What is the purpose of lipofectamine?
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.
What is FuGene 6 transfection reagent protocol?
FuGENE® 6 Transfection Reagent Protocol. FuGENE® 6 Transfection Reagent is a nonliposomal reagent that transfects DNA into a wide variety of cell lines with high efficiency and low toxicity. The protocol does not require removal of serum or culture medium and does not require washing or changing of medium after introducing the reagent/DNA complex.
Can FuGene® transfection be reversed?
Reverse transfections are rapid, convenient and easily automated, and are frequently used for high-throughput experiments. We tested the ability of FuGENE® 6 and FuGENE® HD to “reverse” transfect three cell lines and monitored efficiency and toxicity using the ONE-Glo™ + Tox Luciferase Reporter and Cell Viability Assay.
Which FuGene reagent produces the most luminescence?
Using HEK293 cells, FuGENE® HD reagent produced the most luminescence, indicating the largest production of luciferase (Figure 2). The optimal transfection reagent:DNA ratio for FuGENE® HD appears to be 3:1 with an ideal volume of 10µl. FuGENE® 6 also produced a large amount of luminescence and was greater than that produced by Trans IT®-LT1.