[Zbrafish] Re: dying embryos

Tony Cheuk-Chung Ho via zbrafish%40net.bio.net (by cheuk.chung from gmail.com)
Thu Apr 3 20:58:34 EST 2008


Jim,

I have had experience injecting RNA into zebrafish embryos, and I
think the water is a legit suspect. I would also look into the age of
the fish used in producing the embryos and the pore size of the
injection electrodes.

In my experiments, I have noted that adult fish, that had been
spawning (once a week) for nine months to a year or so, produced
clutches that just die at gastrulation. This observation started out
with at least half the clutch dying, and then rapidly deteriorated to
~70% of the clutch dying without any manipulation.

Anther blunter in my injection experiments is the pore size on
injection electrode, I used to have a batch of pretty big pored
needle(~20um), they just trashed the injected embryos. I now use
electrode with a pore size of about 7um and they work beautifully.

Hope it helps. Good luck with the fish.

Tony

On Apr 4, 2:30 am, "Jim D." <jimdowlingmd... from mac.com> wrote:
> Hello.  We have recently had a problem.  In our last several
> injections of either RNA or cDNA, every embryo has been dying.  They
> look OK at 8 hpf but are dead by 24 hpf.  At least one of the
> constructs has been tried previous and is not lethal by itself.  One
> thought we had is that we are using a batch of Qiagen RNase free water
> for the dilutions.  Otherwise, we are looking for thoughts/
> suggestions.  Has this all of sudden happened to anyone else?
> Thanks.



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