RNA isolation

Tue Nov 15 11:59:14 EST 1994

I favor the method of Chomczynski and Sacchi (1987, Analytical Biochemistry, 
162: 156-159) for isolation of RNA isolation from all stages of zebrafish 
embryos. It's fast, reliable, and gives good quality RNA. If you plan to do 
RT-PCR, you should be careful that the acid phenol extraction removes all 
chromosomal DNA. You may have to treat your RNA with a RNase-free DNase. 
I've never done this on eggs, so I don't know yield. However, from 3 hr old 
embryos, I've recovered 160 ng total RNA per embryo.  I've also used a kit 
from Clontech, the Micro-Scale RNA Separator. It's the same method as 
described above.

Let me know if you have more questions. 
Bruce Appel
Inst. of Neuroscience
University of Oregon
appel at uoneuro.uoregon.edu

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