Transfecting NIH 3T3 cells - looking for effiency for maximum knockdown (Feb/02/2006 )

Hi, I am looking to transfect NIH 3T3 cells with shRNA in plasmid vectors (already made) in order to knockdown/out genes.

I have not worked with these cells before......but I must use a mouse line and this is what someone has available. I am led to believe that transfection efficiency in them is not that high and so am just wondering if anyone can report any successful protocols before I start.

Thank you.

-ajh1269-

QUOTE (ajh1269 @ Feb 2 2006, 07:17 AM)

What transfection method you using? If its something a company provides, maybe they would be able to help you.

-Andtwo-

There are a number of options available, I was just looking for views before I start, the most promising is Amaxa's Nucleofector kit I think but I have never used it.

-ajh1269-

QUOTE (ajh1269 @ Feb 3 2006, 01:01 PM)

There are a number of options available, I was just looking for views before I start, the most promising is Amaxa's Nucleofector kit I think but I have never used it.

I used it once, to transfect old fibroblasts. It worked very well, and I also needed a good transfection yield because I wanted to inhibit some signaling by transient transfection.
Really works well. good assistance. But more expensive than other transfection methods.

-Missele-

Ok thanks for that am hoping to be able to borrow reagents etc for a quick look see then if the results are encouraging I am sure I can persuade someone to spend the money.

-ajh1269-