RNA Extraction from Blood? - Need help with separating classes (Aug/27/2004 )
Hello all. I need some help. I need to isolate RNA from granulocytes, lymphocytes, and monocytes. I am receiving 40cc of whole blood and I am having a tough time separating cell types from each other. I am doing fine with the granulocytes, but the monocyte and lymphocyte pool are mixed. I am using the Histopaque/Percoll procedure to get my fractions. Any other ideas?? I do not have access to a cell sorter. I am using a flow to determine the purity of my samples. Thanks so much.
Monozytes/Macrophages adhere to plastic surfaces, don't they?
So with your granulocytes gone, just incubate the lymphos/macrophages in a plastic vessel for some time. then the lyphos should be still in suspension while the macros should have adhered to the plastic. Scrape them off and there you are!