Lipofectamine RNAiMAX

March 26, 2018

The Pennsylvania State University
Biochemistry and Molecular Biology
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

ThermoFisher Scientific

Product Name:

Lipofectamine RNAiMAX

Catalog Number:

13778075

The goal of the project was to determine if the transcription factor NFIA mediates the cytotoxicity of a novel anti-cancer therapeutic. This product was used to knockdown NFIA in human melanoma cells.

Experimental Design and Results Summary

Application

siRNA transfection reagent

Starting Material

UACC-62 and UACC-257 cells

Protocol Overview

1. Collect and count cells to be plated2. Prepare two tubes: Tube A contains Opti-Mem media and RNAiMAX reagent and Tube B contains Opti-Mem media and siRNA. Amounts of reagent and siRNA can be scaled up or down depending on the number of cells that will be transfected. 3. Add Tube B to Tube A and incubate at room temperature for five minutes. 4. Add Tube A/B mixture directly to cell suspension and plate cells. 5. Cells can be incubated for 1-3 days before analysis. Transfection can also be performed on adherent cells.

Tips

I got great results when I transfected cells while I was plating them. As a bonus, it also saved me time as I didn't have to plate and then wait for the cells to adhere.

Results Summary

An 87% and 90% knockdown of NFIA was observed in UACC-62 and UACC-257 cells, respectively.

DOI or PMID #

None

Additional Notes

N/A

Image Gallery

Summary

The Good

The protocol is clear and easy to follow, especially if one is familiar with other Lipofectamin products (ex. Lipofectamine3000). The protocal can be scaled up or down easily for varying cell numbers with efficient knockdown observed. Cells can be transfected with siRNA either after adherence or during plating.

The Bad

I haven't had any issues yet. Works much better than other siRNA delivery systems I have tried.

The Bottom Line

If you are going to be doing knockdowns, this is a great delivery system to try. Works very well and is easy to use.

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