jetPRIME Transfection Reagent

February 02, 2017

Touro University-CA
Basic Sciences, College of Osteopathic Medicine
Professor

Overall

Quality of Results

Ease-of-Optimization

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Company:

Polyplus

Product Name:

jetPRIME™ DNA and siRNA Transfection Reagent

Catalog Number:

114-01

jetPRIME transfection reagent was tested on DU145 cells and GFP expression was assessed after 24 hours. I estimate a transfection efficiency of at least 50% and as high as 75% from this quick trial.

Experimental Design and Results Summary

Application

DNA transfection into adherent cell

Starting Material

DU145 cells

Protocol Overview

Fifty thousand DU145 cells were plated overnight in duplicate wells of a 24 well dish. Cells were transfected the following day using jetPRIME and 0.5 ug plasmid DNA, according to mfr's instructions. Briefly, DNA was mixed with jetPrime buffer solution, to which jetPrime reagent was added. After a 10 min incubation, the transfection mixture was added directly to the medium in each well. Media was replaced after five hours. Cells transfected with GFP/pCS2 plasmid were visualized about 18 hours later using a fluorescent microscope. In Figure A, cells expressing GFP. In Figure B, using higher brightness, live and dead cells are seen; the number of non-transfected: transfected cells can be seen in this view.

Tips

The product is rather foolproof. I accidentally mixed the DNA/buffer/reagent but forgot that I has 2x wells to transfect, but only used 1x worth of buffer. After adding extra buffer, the transfection results were still accaptable for a trial run.

Results Summary

The transfection efficiency was very good (50-75%) but there was significant cytotoxicity. Each well still had enough cells in it that at the 48 hour mark, I had enough for a western, or an IFF experiment.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Rapid transfection (10 mins worth of mixing); easiest protocol I've ever seen, with suggested amounts of cells to use per vessel, as well as amounts of buffer, DNA and reagent. Good transfection efficiency

The Bad

Significant cytotoxic effect, even with even vector. Buffer alone is not toxic

The Bottom Line

A very easy and rapid protocol

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