microinjection of oocytes - (Nov/23/2006 )

Hi,
I'm trying to inject GV mouse oocytes with a CMV-GFP construct and have a few questions for the "pros". Does the age of the mouse from which you collect the oocytes matter? Is it best to keep the oocytes arrested after injection of the plasmid or can you let them mature to MII? Is a CMV promoter suitable to drive transcription of the plasmid in an oocyte?

Any comments,tip or advice would be greatly appreciated.

Thanks.

I've done the same experiment with great results.

I don't know if the age of the mouse matters, I used 18 day old mice.

You should keep the oocytes arrested after injection. When I examined the fluorescence the ones that had undergone GVBD did not show fluorescence. I used IBMX and the oocytes were cultured in KSOM media under mineral oil.

I think a CMV promoter would be fine.

Let me know if you would like more details.