transfection or injection of siRNA - Which one is better? (Jan/26/2006 )
Hi, dear all,
Can anyone can make me sense that which one can get a better RNAi result, transfection or injection? same siRNA same cell line.
injection??? do you mean electroporation?
that depends. sometimes cells are too fragile to support electroporation. In other hand, some reagents show too much toxcicity for certain cells.
You may tell more of your cell line for having tips on it.
i use lipid mediated delivery and goes well.
I mean, injection is to inject siRNA into cell directly, and cell line is nerve cells. I selectively inject some cells and further culture them. There aren't problem for me because I am good at micro-injection of cells.
Oh, that's interesting. How many cells do you need to inject? One by one?
Yeah, I inject cell one by one, maybe I can inject 150 cells/per hours. Then continue cell culture.
I mean inject cell directly.
For cells refractory to transfection such as primary cells, it certainly worth the effort to do direct injections, but for cells that are easier to tansfect, I don't think so.
Hi . pcrman:
Thanks for your suggestion. I am a beginer in RNAi field and just want to use injection technique to study RNAi.
I also have another question to ask you. which is that, if the target gene of RNAi knockdown isn't expressed in cell culture, just in the brain of the living animal and at protein level, how can I test the expression of RNAi? And how can select a better siRNA from all candidate si RNAs more easily and more inexpensive?
max [font=Comic Sans Ms][size=7]
This is off-topic but anyways....pcrman what transfection kit do you use?? And what cell lines do you work with?
Mostly I use Lipofectamine 2000 and different cancer cell lines and 293 cells.