Reference gene - (Oct/29/2004 )

Hi,

I've been trying to treat cell lines with AZA and TSA and investigate whether expression is increased after treatment. The problem is that I can't find a stable reference gene that isn't affected by the treatment. Does anyone know of a good gene to use or has any ideas?

Thanks in advance

-molbio-

Aza and TSA are epigenetic reagent and may affect genes genome wide. They may also act through other pathways. So reference genes usually used for RT-PCR may also be affected such as beta-actin. I think using 18s and 28s rRNA gene as the control gene is OK.

-rassen-

I have tried S28. At low aza concentrations (1 uM for 96 hrs) it is unchanged but changes expression when treated with 10 uM aza in my cells.

-MDR-

Hi
We treat B cells with Aza and TPA and have used GUS successfully as a reference gene.

Good Luck

-Cilla_-