Posted 21 April 2009 - 09:05 PM
I wanted to know Which anti-coagulant and its working concentration used to separate PBMC and Plasma. I need to use PBMC's for culture and cytokine responce. Plasma to measure cytokine profile using CBA (cytometric Bead Array) as well as reverse Transcriptase PCR from the RNA extracted from the plasma. please can anyone suggest me the anti-coagulant and its concentration to used for separation of plasma and cells.
Posted 22 April 2009 - 05:08 PM
Posted 22 April 2009 - 08:30 PM
waiting for suggestions.
Posted 22 April 2009 - 11:32 PM
Not sure about EDTA and PCR but should not be a problem with Heparin I guess. U can run a control just to be sure. What procedure will you be using to isolate PBMC?
Posted 23 April 2009 - 04:58 PM
Posted 23 April 2009 - 07:05 PM
This is new for me. I am sorry that I mentioned Heparin here though I had mentioned I use it for immunocytometry studies. I googled for Heparin-PCR and found a few studies. Picked up these for reference.
- Effects of heparin on polymerase chain reaction for blood white cells.
- EDTA Is a Better Anticoagulant than Heparin or Citrate for Delayed Blood Processing for Plasma DNA Analysis
- Direct PCR from Whole Blood
It would still be better to run a control to be on safe side whatever U might use.