Is there a place where I can find references from other researchers who have used your reagents?

Product FAQ

Answer

Visit the product page for each reagent type and you will see a list of references at the bottom of the page. A table that lists specific cell line references is also accessible. We also recommend www.highwire.org as a search engine to find a large selection of up-to-date research articles using our transfection products. Simply include the name of the transfection reagent and your cell line/application of interest in your search criteria.

Answer Id: E8968

Was this answer helpful?

Yes
No
Thank you for your response

What recommendations do you have for selecting a transfection reagent?

Product FAQ

Answer

Please refer to the Transfection reagent selection guide (http://www.thermofisher.com/us/en/home/life-science/cell-culture/transfection/transfection-reagent-application-table.html) to make the right choice.

Answer Id: E8961

Was this answer helpful?

Yes
No
Thank you for your response

Is there a place where I can find references from other researchers who have used your transfection reagents?

Product FAQ

Answer

Visit the product page for each reagent type and you will see a list of references at the bottom of the page. A table that lists specific cell line references is also accessible. We also recommend www.highwire.org as a search engine to find a large selection of up-to-date research articles using our transfection products. Simply include the name of the transfection reagent and your cell line/application of interest in your search criteria.

Answer Id: E8999

Was this answer helpful?

Yes
No
Thank you for your response

Why do I see precipitates on the cells after transfection?

Product FAQ

Answer

A small granular-like precipitate may be detected microscopically on the cells after transfection using cationic lipid. This is normal. The presence or absence of this precipitate is not indicative of the transfection efficiency. The precipitate can be caused by presence of excess EDTA or cationic lipid. Use DNA in water or, if in TE, use EDTA concentrations of <0.3 mM in the diluted DNA. Ensure concentrations of cationic lipid reagents do not exceed recommended amounts in complex formation.

Answer Id: E3996

Was this answer helpful?

Yes
No
Thank you for your response

Are lipid transfection reagents fluorescent?

Product FAQ

Answer

The intrinsic fluorescence of Lipofectamine™ 2000 and Lipofectamine™ under FITC conditions was examined and the results were as follows:
- Lipofectamine™ 2000 in serum-free conditions (with or without DNA) - weak whitish fluorescence.
- Lipofectamine™ 2000 in serum-containing conditions (with or without DNA) - medium white and red fluorescence.
- Lipofectamine™ & Lipofectamine™ PLUS (with or without DNA) - orange fluorescence.
- We haven't seen any significant green fluorescence attributable to the Lipofectamine™.

These observations were made after transfection of CHO-K1 cells and by examining the live cells in PBS. Other lipid transfection reagents were not studied extensively on this issue, but it is very likely they will produce fluorescence in transfected cells.

Fluorescence from various media components may be where the problem lies. For example, riboflavin apparently fluoresces in the green wavelength and thus DMEM, which has a high riboflavin content, can be problematic in fluorescence experiments. Ham's F12 medium is 10x lower in riboflavin and is more suitable for fluorescence work.

One possible solution is to perform fluorescence imaging in PBS, rather than in culture medium. Also, check to make sure the cells are healthy and intact. Lysed cells or cells under stress may generate autofluorescent products.

Answer Id: E4451

Was this answer helpful?

Yes
No
Thank you for your response

Do mammalian cells in culture divide in the presence of G418?

Product FAQ

Answer

Yes, cells can continue to divide up to two times in the presence of lethal doses of G418. The effect of the drug usually becomes apparent by two days.

Answer Id: E8969

Was this answer helpful?

Yes
No
Thank you for your response

Are cell density (% confluency) and passage number important considerations for transfection?

Product FAQ

Answer

Yes. Cell density will affect transfection performance. Lipofectamine™ 3000, Lipofectamine™ 2000, and Lipofectamine™ LTX/PLUS provide excellent transfection performance at confluencies between 70 and 90%, while some toxicity may be observed at confluencies lower than this. Lipofectamine™ RNAiMAX works best at confluencies between 60 and 80%.

Passage number may affect transfection experiments. We recommend consistent splitting and plating of cells. Excessive numbers of passages may decrease transfection performance. We do not recommend splitting cells for more than 20-30 passages. If transfection performance declines and cells have been in culture for a long time or excessively/improperly passaged, we recommend that you restart your cultures with a new vial of cells from liquid nitrogen. Please refer to the Gibco™ Cell Culture Basics handbook (https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics.html) for proper guidelines for culturing and passaging cells.

Answer Id: E8962

Was this answer helpful?

Yes
No
Thank you for your response

Which transfection reagent do you recommend for RNAi applications?

Product FAQ

Answer

We recommend using Lipofectamine™ RNAiMAX Reagent for delivery of siRNA into all cell types. It has been specifically developed for siRNA transfection while providing high transfection efficiency with minimal cytotoxicity. As a result, less optimization is necessary. For vector DNA–based RNAi applications, we recommend Lipofectamine™ 3000 Reagent with the P3000™ Enhancer Reagent.

Answer Id: E9000

Was this answer helpful?

Yes
No
Thank you for your response

I've mistakenly frozen my lipid reagent-can I still use it?

Product FAQ

Answer

Freezing can alter the integrity and composition of the lipid particles. The performance of the reagent may be affected, and it may only work for the first week or so. It is safer to purchase a new vial to ensure function.

Answer Id: E9064

Was this answer helpful?

Yes
No
Thank you for your response

Can you compare and contrast the various cationic lipid-based transfection reagents you offer in terms of composition and application?

Product FAQ

Answer

- Lipofectamine™ 3000 & P3000™ Reagents are of a proprietary formulation.
- Lipofectamine™ 2000 Reagent is a proprietary formulation.
- Lipofectamine™ MessengerMAX Reagent is a proprietary formulation.
- Lipofectamine™ RNAiMAX Reagent is a proprietary formulation.
- Lipofectamine™ LTX Reagent is a proprietary formulation.
- Lipofectin™ Reagent is a 1:1 (w/w) formulation of N-[1-(2,3-dioleyloxy) propyl]-N,N,N-trimethylammonium chloride (DOTMA) and the neutral lipid dioleolyphosphatidylethanolamine (DOPE).
- Lipofectamine™ Reagent is a 3:1 (w/w) formulation of the polycationic lipid 2.3-dioleyloxy-N(2(sperminecarboxamido)ethyl)-N, N-dimethyl- 1-propanaminium trifluoroacetate (DOSPA), and DOPE.
- PLUS™ Reagent is a proprietary formulation.
- DMRIE-C Reagent is a 1:1 (M/M) liposome formulation of the cationic lipid DMRIE (1,2-dimyristyloxypropyl-3-dimethyl-hydroxy ethyl ammonium bromide) and cholesterol.
- Cellfectin™ II Reagent is a proprietary formulation.

- Lipofectamine™ 3000 Reagent results in the highest efficiency of plasmid DNA delivery into most cell types. It also works well for siRNA transfection and co-transfection of plasmid DNA with siRNA.
- Lipofectamine™ 2000 Reagent works well with most adherent cells as well as difficult-to-transfect cells. It can be used for transfection of plasmid DNA, siRNA, and co-transfection of plasmid DNA with siRNA.
- Lipofectamine™ MessengerMAX Reagent is recommended for transfection of mRNA or short oligos.
- Lipofectamine™ RNAiMAX provides the best results when transfecting siRNA/miRNA.
- Cellfectin™ II Reagent is recommended for transfection of insect cells.

Answer Id: E9005

Was this answer helpful?

Yes
No
Thank you for your response

Is my lipid (Lipofectin™, Lipofectamine™, DMRIE-C, Lipofectamine™ 2000, Oligofectamine™, Cellfectin™ II) supposed to be cloudy?

Product FAQ

Answer

It is normal to see some turbidity and cloudiness in DMRIE-C, Cellfectin™ II and Lipofectamine™ 2000, and the others should be clear. Lipid transfection reagents are sensitive to low temperature; if you put them at temperature below 4 °C they will precipitate or even freeze and lower the efficiency. Most lipids will go cloudy and precipitate upon freezing, and may not be active anymore.

Answer Id: E4000

Was this answer helpful?

Yes
No
Thank you for your response

Can I use the same amount of any transfection reagent for different cell lines?

Product FAQ

Answer

No. The transfection efficiency is highly dependent on the amount of reagent used per well and may be different between reagents. Please consult the product information that is provided with the transfection reagent for optimal use. The protocol that is supplied with the product will provide you with an optimal range of transfection reagent to use per well. During product development, this range was determined to work well across a variety of cell lines. If you are still not achieving the performance you desire in your particular cell line, further optimization may be necessary.

Answer Id: E8963

Was this answer helpful?

Yes
No
Thank you for your response

Can I use Lipofectamine™ RNAiMAX to co-transfect siRNA with plasmid DNA?

Product FAQ

Answer

While Lipofectamine™ 3000 or Lipofectamine™ 2000 can be used for co-transfection of siRNA with plasmid DNA, Lipofectamine™ RNAiMAX cannot be used.

Answer Id: E9039

Was this answer helpful?

Yes
No
Thank you for your response

I accidentallly left my lipid reagent at room temperature. Can I still use it?

Product FAQ

Answer

Yes, all of our lipid transfection reagents are stable at room temperature for months.

Answer Id: E9065

Was this answer helpful?

Yes
No
Thank you for your response

What is the main advantage of viral transduction over transfection?

Product FAQ

Answer

Transfection does not work for certain cell types such as non-dividing cells, whereas viral transduction works for dividing as well as non-dividing cells, such as neuronal cells that are hard to transfect.

Answer Id: E8979

Was this answer helpful?

Yes
No
Thank you for your response