Hello everyone,
I have been transfecting my primary hippocampal neurons at DIV7. I am just curious to know when and how does one decides what is the best time period to transfect them?
How are you transfecting and what with? We've had best results using lentivirus early (DIV2-3) which seems to kill fewer cells than if we wait longer.
What kind of nucleic acid are you transfecting? If you're using Optifect (which is a lipid based transfection reagent) the transfection is likely to be transient and last for only a few days. So the optimum time to do this really depends on your experimental design. What are you measuring and how mature do you need the cultured neurons to be in order to do that? Aim to transfect at a time point 2-3 days before your intended measurement as a starting point. You might need to test how long your effect takes to appear and disappear. (Eg, transfecting siRNA will take effect sooner and probably last for a shorter period of time vs. transfecting a plasmid that drives a shRNA cassette.)
- Badges
- Science method