RNA from T-cells

[Volker Blaschke]

Dear fellow researchers,

I need to make RNA from Buffy-coat (native) T-cells. However, with all 
the protocolls I've used so far (e.g. GuSCN/Ultracentrifugation, 
Chomczynski/Sacchi and Qiagen RNeasy kit), yield was incredibly low 
(around 5 ug total RNA from 20 million cells).

Fort Northerns, I need much more than this, but cell supply is obviously 
limited.

Does anyone know if this yield is in fact what one gets from native 
T-cells or of another methods giving higher yields?

Qiagen states in their protocoll that their yield is around 5 ug total 
RNA from 1 million T-cells, but what they don't say (and can't say on 
the phone) is if that's native cells or stimulated cells or a cell line.

Thanks for your help in advance!

  Volker


Dr. Volker Blaschke
Dept. of Biochemistry
AG Prof. Jungermann
Humboldtallee 23
D-37073 Goettingen
Germany

Email: vblasch at gwdg.de
Tel.: xx49-551-39-5959 / -5978
FAX :            -5960