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RNA Extraction from Blood?


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#1 NewTech

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Posted 27 August 2004 - 11:33 AM

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.

#2 jadefalcon

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Posted 30 August 2004 - 01:12 AM

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!

mike

Edited by jadefalcon, 30 August 2004 - 01:12 AM.

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