Hello people! I'm trying to transfect my monocyte-derived dendritic cells(MoDC) with some siRNA using Neon electroporation system. The protocol recommended to use buffer T for primary blood-derived suspension cells and buffer R for primary adherent cells. I would think that my MoDCs belong to "primary blood-derived suspension cells" because they have been sitting in culture for 4 days before the transfection, but in a paper by Sprokholt et al (Article RIG-I–like Receptor Triggering by Dengue Virus Drives Dendri...
) they used buffer R for both cultured and primary dendritic cells.So I wonder if anyone knows the actual difference between these two buffers? Why is one better than another for different cells?
For the proprietary buffers found in neon transfection kits
Resuspension buffer R: iPSC and majority of adherent cell lines
Resuspension buffer T: blood derived suspension cells only
Electrolytic buffer E: used with 10ul neon tips
Electrolytic buffer E2: used with 100ul neon tips
Resuspension buffer R is the standard cell resuspension buffer. It is used to resuspend established adherent and suspension cells as well as primary adherent cells. Maximum voltage is 1900.
Resuspension Buffer T is an alternative that is used to resuspend primary T and B cells, PBMCs, monocytes, and bone marrow–derived cells. Its composition differs from that of Buffer R (thus allows for higher voltage) and it doesn't work with established cell lines or primary cells, which have been kept in culture for some time.
If you are unsure which would work, manufacturer recommends testing both in separate optimization experiments.
for more details refer to
-https://www.youtube.com/watch?v=lvRbFyQ_m3g (21:31)
-https://www.thermofisher.com/ng/en/home/technical-resources/technical-reference-library/transfection-support-center/electroporation-support/electroporation-support-getting-started.html
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