RNA isolation from zebrafish embryos
Kathleen.Kayes-Wandover at UTSouthwestern.edu
Mon Aug 6 16:42:20 EST 2001
I'm a researcher in Pediatric Endocrinology at the University of
Texas Southwestern Medical Center in Dallas. We're studying a
novel G-protein coupled receptor which is found in zebrafish &
would like to study it's role in development. I'm interested in
isolation of RNA from zebrafish embryos for RT-PCR and RACE. I
would like to use the UltraSpec-II RNA isolation system from
Biotecx (this is what I generally use). If you're not familiar with this
reagent, it involves homogenization of the sample with RNA
extraction reagent (a mix of guanidine, urea, phenol,etc) and
extraction with chloroform. The aqueous phase obtained after
centrifugation (12,000g for 15 minutes) is mixed with a resin which
binds the RNA & then is precipated with isopropanol.
Two questions: (1) is this method appropriate to use (2) do I need
to remove the chorions, if so, how would I do it ?
Thank you for your consideration.
Kathleen M. Kayes-Wandover, Ph.D.
UT Southwestern Medical Center at Dallas
email: Kathleen.Kayes-Wandover at UTSouthwestern.edu
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