RNA from cultured Osteoblasts - osteoblasts (May/14/2004 )

We have been trying to extract RNA from cultured primary human osteoblasts using Tri Reagent with very poor yields.

We initially tried 6 well plates but have recently been using 100mm Diameter culture dishes.

Do we need bigger dishes?
Is there anything special about Osteoblasts that we've missed?????

Did you scale up the amount of Tri Reagent added to your dishes? since you said you used to use 6-well plate. Stick to what the manual says about the amount of Tri reagent needed. If it is not enough, you will get low yeild of RNA.

Don't you grow these things in calcium free medium?