RNA extraction from splenocytes - (Oct/17/2007 )

Hi all,

I am a new member at the forum.

I am trying to extract total RNA from mouse splenocytes to use it in real-time RT-PCR. The problem is I always get very low quantities of RNA.

The last time, I used Promega's mammalian RNA extraction kit using close to 100*10^6 cells. The RNA concentration I got was only 0.037 ug/ul.

The next time I will try TRIzol reagent, but the thing is that I only have 7 ml of it and so not much room for trial and error. So, can anyone suggest a proper number of cells to use?

I will be grateful for any help. Thanks.

-scientifica-

I used Qiagens RNAeasy kits for mouse splenocytes RNA isolation, with about 1-10 10E6 cells to start with, and that worked fine for me.

mike

-jadefalcon-

Hi mike

Thank you for your reply. Could you please tell me what yield of RNA do you get from 1*10^6 cells?

Regards
Maysaa'

-scientifica-