Hi - I was hoping to optimise nucleofection for PAEC. Has anyone optimised the transfection before? Which of the Amaxa programs works best? A shame that you cannot optimise Amaxa in any other way than using trial & error...
Dear Svenja,
this post may appear a little off topic, but I would like to present an alternative to nucleofection: the magnetofection technology***
In this method, complexes of nucleic acids and magnetic nanoparticles enters the cell via classic endocytosis pathways, and no hole is created into the cell membrane. Thus the viability is not hampered.
Magnetofection is really powerful and ideal for primary cells such as neurons, epithelial cells and of course endothelial cells (and more...) as demonstrated by our citation database (in the link below).
PolyMag Neo would be the reagent of choice;its protocol is really simple: use 1 µL PolyMag Neo per µg DNA; incubate 20 min at RT before adding onto cells. place the cells onto the magnetic plate and that's it.
would you like to try it as an alternative to nucleofection, please do not hesitate to contact me via ResearchGate or directly at [email protected]
Best regards,
Cedric
MAGNETOFECTION
*** The Magnetofection technology uses a magnetic field to attract and concentrate complexes of magnetic nanoparticles and nucleic acid onto the cell surface as demonstrated by Grześkowiak BF et al, Pharm Res. 2014 Jul 18. (http://www.ncbi.nlm.nih.gov/pubmed/25033763).
http://www.ozbiosciences.com/module/citationfinder/default
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