RNA ext. from vessel endothelial cells

QIAGEN QIAGEN at kaiwan.com
Tue Jun 20 11:45:07 EST 1995

In Message-ID: <glarson-090695173836 at>
glarson at ccmail.llu.edu (Garry P. Larson) wrote:

>Can any1 give me a good protocol for preparing RNA from arterial
>endothelial cells directly from mammals?  The endothelial cells comprise
>perhaps only 10% of the total cell population compared to connective tissue
>cells.  Can one just use a Qiagen RNA extraction kit (or similar company
>product) after grinding the heck out of the tissue?  Or is there a better
>way?  Do you have any protocols? I'd sure like some good recommendations.

I checked whether we have any references of customers using QIAGEN 
RNA extraction kits (RNeasy) specifically for arterial endothelial cells 
directly from mammals, but I did not find any. RNeasy has been used for a 
wide variety of tissues, including brain, heart, intestine, kidney, liver, 
muscle, lung, spleen, bone material and some more unusual samples such as 
juvenile lobster claws. 

The procedure should work as long as the homogenization is efficient Ñ 
"grinding the heck out of the tissue" is one option, using a 
rotor/stator homogenizer such as Polytron from Brinkmann is another.
If the lysate is still very viscous, increasing the buffer volumes 
and using a syringe with needle to shear the high molecular 
weight DNA usually helps.
If you have any questions, please do not hesitate to call me or our tech 
service department at 800-426-8157.

Brigitte Steude Masone
Product Manager
QIAGEN       800-362-7737
Temporary Email Address          qiagen at kaiwan.com

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