Are there anyone who performed monocyte ısolation from peripheral blood by using The EasySep™ Direct Human Monocyte Isolation Kit? In my research project i want to examine the expression of a Mirna and mrna in monocytes. I'm curious about the volume of peripheral blood needed for obtaining good quality and quantity RNA.
A good amount of RNA to start with is 1µg and 1µg RNA can be generally isolated from 10^6 monocytes. Since monocytes roughly represent 10% of PBMCs you would need about 10.10^6 PBMCs to have enough material. The number of PBMCs per volume of blood can vary from one person to another but using 20 mL you should be relatively comfortable.
The other way around, 20 mL blood should be generally enough to isolate good quality RNA from isolated monocytes.
Thanks for your answer. Did you use which kit for this protocol above?
I think you should do a standardization run with how much monocytes (percentage) and pbmcs (count) you are getting in a standard volume of blood (like 1ml or 100 microlitre) and then calculate how much blood you need. Using someone else's protocol incurs a bit of risk. However I have not used the monotype isolation kit.
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