One of my students discovered these worms in our zebra tank this morning. There were ~ 30 of them along with some other organisms. The fish seem healthy and are breeding fine, but I want to get rid of them. Any ideas?Attachment 423

Attached Images

worms.jpg (63.0 KB)

Dear Wei,

Very likely your problem is due to the slow maturation of DsRed.
mCherry should be better and is fairly bright. We have also had good
success with TagRFP, which is extremely bright (available commercially
from Evrogen). Another possibility is tdTomato, which is extremely
bright. We have not used it ourselves but have heard good things from
others. Don't know about maturation time.

There is an easy experiment to test whether maturation time is the
issue: do a DsRed in situ on your transgenics, and see if the
timecourse mimics the native gene.

best wishes,
Chi-Bin Chien

On Nov 2, 2009, at 5:19 PM, Wei Xia wrote:

Has anyone encountered low 260/230 readings (<0.2) using the Trizol
RNA extraction. We're getting okay yields and great purity using 5
embryos (24hpf-72hpf)/sample, but are having problems with what we
assume is contamination.

We have already tried a number of things: bought all our chemicals
new, repeat chloroform extraction 2 times, do EtOH cleaning step 3
times. We homogenize manually with pestles.

Thanks for any advice.

Our conductivity is routinely between 500-700 and we don't have an issue with this.



Hope this helps,

Becky



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From: zbrafish-bounces@oat.bio.indiana.edu [mailto:zbrafish-bounces@oat.bio.indiana.edu] On Behalf Of Evert-Jan van den Brandhof
Sent: Tuesday, November 03, 2009 1:49 AM
To: zbrafish@magpie.bio.indiana.edu
Subject: [Zbrafish] zebrafish gill disease?




Hello,

Is anyone familiar with (what looks as) fast growing epthelium on upper gill covers?
We do'nt have any problem with egg production but it may cause some stress on breathing capacity?

see picture:

could anyone reconfirm this could be caused by conductivity?

system: Zebtec (http://www.tecniplast.it/_assets/pro...G_Internet.PDF) <http://www.tecniplast.it/_assets/products/CAT012_ING_Internet.PDF>

pH= 7.5
Conductivity= 500 µS
Temperature= 27,5 °C

Conductivity and pH are automatically adjusted with Seesalt and Sodiumbicarbonate.

Thanks in advance,

Evert-Jan


¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤
Evert-Jan van den Brandhof, BSc
National Institute for Public Health and Environment (RIVM)
Laboratory for Ecological Risk Assessment (LER)
P.O. box 1, 3720 BA Bilthoven, The Netherlands
Tel: (+31)30-2743544
http://www.rivm.nl/milieuportaal/ <http://www.rivm.nl/milieuportaal/>
¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤



Disclaimer RIVM <http://www.rivm.nl/disclaimer.htm>

Dear All,
We are wondering about what kinds of red fluorescent reporters are
good for making transgenic zebrafish.
Our gene’s strongest expression occurs around 24hpf by in situ
results. But when we used DsRed as the reporter we could not see any
fluorescence until after 44-48hpf in our transgenic embryos. The
fluorescence was pretty obvious at 24hpf if transient expression was
checked by plasmid injection. So we guess the late detection of DsRed
in these transgenic embryos is not because of the promoter we used is
not complete or something. We thought this is might because of DsRed’s
long maturation half time. We are thinking about using monomeric
mCherry but worry about its low brightness compared to DsRed.
If you have any experiences of making RFP transgenic fish please share
them with us!
Thanks in advance for your testimonies on this.
Wei