problems extracting RNA from dendritic cells

[ateijeiras]

I'm getting very low amounts of RNA when extracting it with TRIzol form under culture differenciated human dendritic cells. I usually don't get more than half a microgram. Could somebody help me or has have the same experience?

[CharonY]

There are a lot of potential reasons, most notably the presence of RNAses. What precautions do you take? Are all you extractions low or only with dendritic cells?

[ateijeiras]

i just have problems with DCs, probably they have a lot of RNAse activity and low amounts of mRNA as they are not very active. I just want to know if somebody have met the same problem with this cells or similar cells, perhaps of the immune system

Thank you for your reply

[CharonY]

Well, I have not used dendritic cells myself, but with older, almost apoptotic cell cultures I used to get far lower yields, too (roughly half of active cells). But absolute quantities are hard compare, of course, at least if not normalized to cell numbers.