Hi,
I am performing transfection studies with mRNA.
I have available Lipofectamine (expired 3 years ago), RNAiMAX (expired 3 years ago) and two recently bought tubes of MessengerMax which were accidentally stored in the freezer for approx 1 week.
Has anybody used RNAiMAX for transfection of mRNA and got good transfection efficiency?
Have you used one of these lipofection agents once expired or after being stored in the freezer for some days?
Which of the options available would you recommend to use? We just bought the tubes of MessengerMax which were frozen so we would like to avoid buying them again (it is very expensive).
Thank you very much!
Sandra
Hi, I have used Lipofectamine 2000 (expired 2019, stored at 4 degree the whole time) for sgRNA transfection into epithelial cells. The transfection has worked well if it helps!
Hey Sandra,
we struggled for a long time to transfect macrophages.
Viromer substances or Lipofectamine either did not work or killed the cells. Moreover, every attempt to transfect DNA led to macrophage death and nothing was expressed.
However, we recently established a method for transfection with the JetMessenger transfection reagent from Polyplus and in vitro transcribed and immunologically silenced mRNA.
For a detailed protocol (including GFP as an example) please have a look at:
Article Highly Efficient Transfection of Primary Macrophages with In...
It worked for peritoneal macrophages, BMDM and MEFS with transfection efficancies between 65% (PMs) and 90% (BMDM and MEFs). But it only delivers mRNA and not plasmids. Tissues macropages go instantly into apoptosis, when we tried to transfect DNA.
I hope this was helpful to you.
All the best and stay healthy,
Marc
Both of them should still work well. Certainly, the type of cells is very important.
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