How long can you incubate your cells with transfection reagent DNA incubated with Lipofectamine?
Add the 500 μl of DNA-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 24-72 hours until they are ready to assay for transgene expression.
How do you store Lipofectamine?
Contents & Storage Contains one vial (1.5 ml) Lipofectamine™ 2000 Reagent. Store at 2–8°C. Do not freeze.
What is the role of lipofectamine in 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 long can Transient transfection last?
24 to 96 hours
Depending on the construct used, transiently expressed transgene can generally be detected for 1 to 7 days, but transiently transfected cells are typically harvested 24 to 96 hours post-transfection.
Should I change media after transfection?
A complete media change can be performed 5 – 24 hours after transfection for very sensitive cells. For most cells, we recommend the media be changed only at 48 hours post-transfection until protocol optimization requires this extra media change. A complete media change should be performed at 48 hours post-transfection.
How long can you store Lipofectamine?
Contents & Storage Store at 2–8°C. Lipofectamine™ Transfection Reagent is guaranteed stable for 6 months when stored properly.
Is Lipofectamine toxic to cells?
Although Lipofectin showed the lowest toxicity to HepG2 cells (89.54% viability), it displayed the lowest transfection efficacy too (8.29%). Generally, HepG2 cells displayed resistance to the toxicity of majority of the reagents tested, with Lipofectamine 3000 (70.59 % viability), being the most toxic reagent.
How long does Stable transfection last?
Depending on the cell type, drug-resistant clones will appear in 2–5 weeks. Cell death should occur after 3–9 days in cultures transfected with the negative control plasmid.
What is the transfection protocol of Invitrogen Lipofectamine 3000?
Transfection protocol On the day of transfection, which should be 1 day following cell plating, perform the following steps, which have been optimized for a single well of a 24-well plate using Invitrogen™Lipofectamine 3000 Transfection Reagent: Step Tube Complexation components Amount per well (24-well)
Why choose Lipofectamine® transfection reagent?
When used in combination with PLUS™ Reagent, Lipofectamine® Transfection Reagent has been shown to work well with cells such as BHK-21, NIH 3T3, COS-1, fibroblasts, keratinocytes, HT-29, MRC-5, and SK-BR3. With such a legacy comes a commitment to providing the best reagents for all scientists and labs, regardless of their funding capabilities.
What is the difference between FuGene HD and Lipofectamine 3000?
Lipofectamine 3000 reagent provided higher GFP transfection efficiency than Lipofectamine 2000 and FuGENE HD reagents for all five cell lines. Figure 2. The transfection protocol for Lipofectamine 3000 was developed to be easy to use while still ensuring optimum performance and reliability in a wide panel of cell lines.
What is the best protocol for cell-specific transfection?
Cell-Specific Transfection Protocols NOTE: Invitrogen Lipofectamine Transfection Reagent Protocols have been optimized for efficiency, viability, and reproducibility across a broad range of cell types. This is often the best place to start, especially in a new cell line.