Reference gene - (Oct/29/2004 )
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
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.
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.
We treat B cells with Aza and TPA and have used GUS successfully as a reference gene.