summary of whole blood preservatives for sorting and mRNA works

From: Fang-yao Stephen HOU (shou@uci.edu)
Date: Fri Dec 21 2001 - 20:19:13 EST


Hello,

I am posting the summay of the info I received, regarding the search of
the whole blood preservatives for sorting and mRNA works. Thank you for
those who responded to my inquiry. Although there isn't any perfect
solution yet, I hope the info here will provide leads in the future.

========
Currently two such products exist, one by Ambion and the other by Qiagen,
both work off the same principal being that they neutralize Rnases and all
other proteins in the blood.  I just was informed about eh Qiagen product
by a rep last week, so its new to the market. Qiagens product is one where
the blood is directly drawn into the collection tube along with the RNA
degradation inhibitor already present so real time RNA levels are
qmaintained.

PAXgene Blood RNA System. You may wish to visit the site at
http://www.qiagen.com/catalog/chapter_06/paxbloodrnasystem.asp
It seems to be very suitable for this application. However, it will lyse
the cells according to the conversation with the technical support people.
Thus, sorting of some subsets may not be possible. It is good for
maintaining mRNA level in the whole blood.

I can not locate the product by Ambion.


===========
I don't have a certain answer for your mRNA question, but I advise trying
Proclin before you give up.  It's a metabolic inhibitor we used (back in
the days when I worked for CompuCyte Corp.) to use to stabilize whole
blood for clinical trials looking at surface markers.  You may still get
some degradation--RNase is EVERYWHERE--but at least it'll shut down
transcriptional changes triggered by activation in transport....

Although through web serach, I found out that ProClin was manufactured by
Rohm and Haas. I can not find the product on the web (www.RohmHass.com)
any more. However, Sigma carries ProClin 150/200/300. Any publication
using ProClin in this application will certainly help.

===========
My experience is that there are good perservatives that stabilize the
blood, but they tend to cross link mRNA and prevent analysis. The samples
work great for flow. Proceed with caution.

=========

Fang-Yao Stephen Hou, Ph.D.
Flow Cytometry Specialist
UC-Irvine



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