What are the two chemical reagents that can be used for transfection?

Transfection reagents can be generally divided into liposomal or high-lipid based and non-liposomal based reagents. Non-liposomal reagents can be mixed protein-lipid reagents, non-liposomal lipids or non-protein non-lipid reagents like dendrimer.

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.

What is the role of Lipofectamine 3000 reagent?

Lipofectamine 3000 reagent leverages our most advanced lipid nanoparticle technology to enable superior transfection performance and reproducible results. It delivers exceptional transfection efficiency into the widest range of difficult-to-transfect and common cell types (Figure 1) with improved cell viability.

How do you add lipofectamine to a cell?

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.

What is transfection explain different methods of transfection?

Transfection methods include various approaches (physical and chemical methods) that are carried out by non-viral techniques – electroporation, calcium phosphate exposure, liposome-based transfection that allow to deliver cargo molecules through cellular membrane without any permanent damage to the cell.

Is Lipofectamine stable transfection?

Yes Lipofec can be used for stable transfection. As mentioned by Michael, anti biotic selection is usualy used. My case is : I got a plasmid in pCDNA3 vector, it contains Neomycin resistance gene and I want to use this plasmid and Neomycin to stably transfect Neuro2A cells using Lipofectamine.

Does Lipofectamine expire?

According to ThermoFisher, lipofectamine 3000 can be used until 2 years after the manufacturing date if stored properly (stored in a 4 degree fridge should be fine). The Optimem is sensitive to expiration. I would recommend using the new reagents for your experiment. Hope this helps!

What is P3000 Enhancer reagent?

P3000 reagent: This reagent will be added to the diluted DNA solution, and its function is to help nucleic acids entering both the cell and the nucleus. For a more detailed description, check molecular mechanism of Lipofectamine 3000.

How does P3000 reagent work?

Lipofectamine 3000, when added to the mixture coats p3000 lipid droplets and confers a positive charge to the outside of the liposhperes. This positive charge allows endocytosis of the lipid droplets into the negatively-charged cell membrane.

Why is optimem used in transfection?

OPTI-MEM is the best option since transfection efficiency is high when cells are grown in O-MEM during transfection. FBS in regular media hinders transfection so using Opti-MEM is good option when using Lipofectamine. N.B: Effectene (qiagen) is a transfection agent optimized to be used with media containing DMEM.