HELP! Problems with RNA extraction from rabbit carotid
jltwong at hotmail.com
Tue Jan 15 06:38:49 EST 2002
I have been trying to extract RNA from cholesterol-fed rabbit carotids with
very variable success.
I initially started with homogenization in a guanidindium thiocyanate and
mercaptoehtanol solution followed by extraction with phenol/chloroform.
Because the protocol took 8 hours and yields were variable, I tried TRIZOL
and Stratagene's Strataprep total RNA Miniprep kit, neither made much
difference - I'm getting a lot of smears.
I normally homogenise my sample by grinding it to powder in liquid nitrogen,
then further homogenising the result in a mechanical Teflon-glass pestle and
mortar. I switched to a homogeniser with a rotating blade, but that didn't
actually break up the rabbit carotid ?because of the high elastin content,
and made little difference to my success rate.
If anyone has got a working protocol for extraction of RNA, or
troubleshooting advice, I'd be very interested!
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