My lipofectamine 2000 accidentally stored in -20 for 5 month after purchasing. Is it potential to still use the reagent? And any recommendation for the optimization? Because I have already transfected plasmid that contain GFP using that lipofectamine and get failure.
Thank you
Freezing Lipofectamine 2000 at –20°C can disrupt its lipid structure, likely reducing transfection efficiency. That said, I wouldn’t discard it just yet — it’s worth testing empirically.
Try using a known positive control and optimize the DNA:Lipofectamine ratio (e.g., 1:2 or 1:3). Use healthy cells at 70–90% confluency and form complexes in serum-free media. If transfection still fails, the reagent is probably compromised and you might have to consider replacing it.
Good luck!
Hello Fami Israyusnita,
Freezing Lipofectamine 2000 at -20 deg C can disrupt the lipid structure leading to aggregation or fusion of the lipid particles, thereby affecting its ability to form complexes with nucleic acids and reducing its ability to efficiently deliver DNA or RNA into cells.
The frozen Lipofectamine 2000 can best be used by thawing it carefully at room temperature, gently mixing it, and then testing its performance in the transfection assay. Compare the results with a freshly opened vial of Lipofectamine 2000 to determine if the frozen reagent is still acceptable for your experiments.
Best,
Dear Fami Israyusnita
I hope you have resolved your issue,
I am jumping in to offer you an alternative to transfect efficiently mRNA; here at OZ Biosciences we develop transfection reagents, mRNA and LNP since more than 20 years and we offer solution ot any kind of transfection.
► Considering mRNA transfection, I would like to present you RmesFect a transfection reagent specifically developed for mRNA delivery. Our technology relies on biodegradable lipids and we use proprietary ones that are very stable and present low index of toxicity.
Indeed RmesFect can be frozen at -20°C without any problem; the preservation is thus better.
→ as a matter of fact, a very recent publication was published last month; https://doi.org/10.12688/verixiv.1800.1.
■ I am sure it could be worth trying it !! ■
If you need any information or if you want to try RmesFect, please do not hesitate to contact me directly at:
I will be glad to send you a sample,
best regards,
Cedric
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