RNA ext. from vessel endothelial cells
QIAGEN at kaiwan.com
Tue Jun 20 11:45:07 EST 1995
In Message-ID: <glarson-090695173836 at 188.8.131.52>
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
Temporary Email Address qiagen at kaiwan.com
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