Lipofectamine 3000-Enhanced Transient Transfection

Johns Hopkins School of Medicine
Oncology
Postdoctoral Fellow

Overall

Quality of Results

Ease-of-Optimization

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Company:

Thermo Fisher Scientific

Product Name:

Lipofectamine 3000

Catalog Number:

L3000-150

Similar to earlier generations, lipo3000 is used to transfect a broad spectrum of cell lines such HEK293T with plasmid DNA. Transfection of cells with plasmid DNA can be optimized based on various parameters, and the procedure is simple and quick. Depending on the cell line, there can be some viability issues associated with the reagents and amount of DNA used. This reagent is very good for efficient transfection.

Experimental Design and Results Summary

Application

Transient DNA Transfection

Starting Material

HEK293T cells are the easiest to test and plasmid DNA

Protocol Overview

The kit can be used by following the manufacturer's protocol. Make sure your cells are in antibiotic-free media to ensure optimal transfection. Plamid DNA is diluted in OptiMEM (invitrogen) and P3000 reagent is added to the diluted at 1:2 ratio. Lipo3000 reagent is diluted in OptiMEM in a separate tube. Finally, the diluted DNA and lipo3000 are mixed and incubated 20 minutes at room temperature for proper lipoplexes/DNA.

Tips

Try various plasmid DNA concentrations to maximize protein expression and cell viability.

Results Summary

We are able to get good transfection in HEK293T cells. Protein expression is quite high in this cell line. We also get transfection in other cell lines such MEFs.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Very good efficient transfection

The Bad

Expensive in the long run so we buy the larger size

The Bottom Line

Highly recommend this reagent for transfections even though the cost is high since we get reliable results.

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