RNA isolation from zebrafish embryos

Kathleen Kayes-Wandover 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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