From naruse from nibb.ac.jp Thu Nov 1 05:27:51 2007 From: naruse from nibb.ac.jp (Kiyoshi Naruse) Date: Thu Nov 1 10:46:07 2007 Subject: [Zbrafish] NIBB International Practical Cource Message-ID: <006201c81c71$db2004d0$b22e3085@Naruse> Dear all, We announce the NIBB International Practical Course on 2nd Developmental Genetics of Zebrafish and Medaka. NIBB International Practical Course 2nd Developmental Genetics of Zebrafish and Medaka Date: March 3rd -12th 2008 Location: National Institute for Basic Biology, Okazaki, Japan Language: English Zebrafish and medaka are excellent vertebrate models for genetic study. Thanks to transgenic technique and embryonic manipulation, these animals are also useful for precise analysis of many events during vertebrate development. The course will cover basic technologies, including gene knockdown by injecting anitisense morpholino-oligo, as well as a number of advanced techniques such as BAC homologous recombination mediated transgenesis, genomic/bioinformatic techniques, and cell tracking using photoconvertible fluorescent proteins. A guest lecturer will discuss emerging topics in the zebrafish and medaka fields. Informal interaction between students and course faculty is encouraged. This is the second international practical course supported by the National Institute for Basic Biology (NIBB) in Japan and is mainly designed for young scientists and those wanting to study in a new field. Travel and accommodation support is available to participants from all countries. Application deadline is November 30th? 2007. For more details see the course website: http://www.nibb.ac.jp/course/index.html Organizing committee Shinji Takada ---------------------------------------- Kiyoshi NARUSE Ph.D. National Institute for Basic Biology, Laboratory of Bioresources, Nishigonaka 38, Myodaiji, Okazaki 444-8585, Aichi, Japan TEL: 0564-55-7581?TEL/FAX: 0564-55-7580 email: naruse@nibb.ac.jp ----------------------------------------------------- From ulrike from mail.nih.gov Thu Nov 1 14:24:40 2007 From: ulrike from mail.nih.gov (ulrike@mail.nih.gov) Date: Thu Nov 1 14:45:52 2007 Subject: [Zbrafish] caMOD 2.3 released - contains customization for zebrafish entries Message-ID: <1193945080.914459.172550@o3g2000hsb.googlegroups.com> The National Cancer Institute's Center for Biomedical Informatics and Information Technology is proud to announce the release of version 2.3 of the Cancer Models Database (caMOD). The new version was released on October 30th, 2007. caMOD 2.3 can be accessed at http://cancermodels.nci.nih.gov/ The cancer models database (caMOD) is a web-based resource that provides information about animal models for human cancer to the public research community. caMOD provides the following key capabilities to its users: - Data Submission--Data in caMOD are extracted from the public scientific literature by curators and verified by the scientists who generated or worked with the models, or they are directly submitted by scientists. - Data Search--Users can retrieve information about the making of models, their genetic descriptions, histopathology, derived cell lines, associated images, carcinogenic interventions, microarray data, and therapeutic trials in which the models were used. caMOD provides links to PubMed for associated publications and other resources such as model repositories, detailed information about altered gene, pathway affected, and information about human clinical trials that utilize the same compounds as the pre-clinical trials in the animal models. - System Function Administration--The Admin function provides services for user registration, review of submitted models and database management. No user account is required to query the database. With this version, caMOD starts customizing the application for models in a variety of species beyond mouse models. caMOD 2.3 supports the submission of zebrafish and rat models. Depending on the selected species, the application uses species-specific vocabularies for anatomy and disease on the Histopathology, Cell Lines, Graft, and Associated Expression pages. Links to the Zebrafish Model Organism Database (http://www.zfin.org) and to the Rat Genome Database (http:// rgd.mcw.edu/) are provided on the Genetic Description pages and allow the user to retrieve information about the modified alleles. Links to the above mentioned resources have also been added to the publication listings. Data about other species can be submitted to caMOD, but no vocabularies for anatomical or diagnostic terms are provided at this time. The Search pages now utilize the species-specific vocabularies. The selection drop down list of model species on the Search pages is created dynamically depending on which models have been approved during the review process. caMOD 2.3 further expands the use of vocabularies provided by NCI Thesaurus (http://nciterms.nci.nih.gov) by introducing the staining method vocabulary to the Image portion of the application. Acknowledgements The rat vocabularies were retrieved from RENI. We would like to thank the members of the Rat Nomenclature Reconciliation Subcommittee for their support and advice. The zebrafish vocabularies were provided by ZFIN, the Zebrafish Model Organism Database, and rendered from bioontology.org. RENI: http://www.item.fraunhofer.de/reni/rat_nomenclature/index.htm ZFIN: http://www.zfin.org Bioontology: http://www.bioontology.org Our special thank you goes to Dr. Hatem Sabaawy who served as the domain expert on zebrafish models. We would also like to thank the NCI CBIIT Enterprise Vocabulary Systems team for their work in converting and importing the vocabularies. The caMOD Team From marcuswrath from starpower.net Fri Nov 2 08:09:57 2007 From: marcuswrath from starpower.net (marcuswrath@starpower.net) Date: Mon Nov 5 12:50:56 2007 Subject: [Zbrafish] 2 positions available in Bethesda, MD Message-ID: <1194008997.553963.255730@d55g2000hsg.googlegroups.com> Aquatics Specialist III Location: Bethesda, Maryland, USA Company: Charles River Laboratories Company URL: http://www.criver.com Salary: $35,000 annually (negotiable) Closing Date: Saturday, December 01, 2007 Date Posted: Thursday, October 25, 2007 Qualifications: High School Diploma or GED is required. A Bachelor degree in biological sciences is preferred. Two or more years experience working with aquatics species is required. Must become AALAS certified at the LATg level within 6 months of eligibility Description: Basic knowledge of aquatic husbandry and technical skills is required and must be capable of identifying sick animals and providing medical treatment as prescribed. Must have knowledge of aquatic species including: zebrafish, sea urchins, frogs, bullfrogs, and any other aquatic or semi-aquatic species that may be utilized in biomedical research. Contact: Contact Name: Matthew Rinker Contact Phone: 240-686-4367 Contact Email: matthew.rinker@crl.com Other Contact Information: Fax 240-686-4392. ----------------------------------------------------------------------------------------------------------------------------------- Aquatics Specialist IV Location: Bethesda, Maryland, USA Company: Charles River Laboratories Company URL: http://www.criver.com Salary: $40,000 annually (negotiable) Closing Date: Saturday, December 01, 2007 Date Posted: Thursday, October 25, 2007 Qualifications: A Bachelor degree in biological sciences is required. Two or more years of experience working with aquatic species is required. Must become AALAS certified at the LATg level within 6 months of eligibility. Description: Provide technical research services directly supporting research protocols and work in partnership with research staff members in the conduct of biomedical research. Provide husbandry and colony management support in multiple facilities for aquatic species such as zebrafish, sea urchins, frogs, bullfrogs, and any other aquatic or semi-aquatic species that may be utilized in biomedical research. Maintain and troubleshoot aquatic support equipment and perform water quality tests. Contact: Contact Name: Matthew Rinker Contact Phone: 240-686-4367 Contact Email: matthew.rinker@crl.com Other Contact Information: Fax 240-686-4392. From scover from pacbell.net Tue Nov 6 05:05:18 2007 From: scover from pacbell.net (Spike Cover) Date: Tue Nov 6 11:55:06 2007 Subject: [Zbrafish] Cloning in Fish--Nucleocytoplasmic Hybrids Message-ID: <47303C5E.7040309@pacbell.net> Dear Ms. Yang Jianhua, I saw an old (1999) post on a forum advertising a book titled, Cloning in Fish--Nucleocytoplasmic Hybrids, by Shaoyi Yan DSc. At that time, it was offered for $28 US plus $8 postage. Is that book still available? If it is written in English and it is available, I would like to order a copy. I hope you can help me order this book. My thanks in advance. Walter Cover Mission Viejo, California USA Reference: http://www.bio.net/bionet/mm/zbrafish/1999-April/004217.html -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20071106/3c6d561e/attachment.html From xiaw from umbi.umd.edu Thu Nov 8 13:33:30 2007 From: xiaw from umbi.umd.edu (wei xia) Date: Thu Nov 8 13:46:43 2007 Subject: [Zbrafish] Help Help with microinjecting RNA Message-ID: <4BEB7629-6442-4A04-A836-5AB1B8DB8985@umbi.umd.edu> Can you please forward my request to the author of this post? Cause I have the same problem that he had met and I really can't figure it out now. Thank you very much for your help. Wei From xiaw from umbi.umd.edu Thu Nov 8 13:46:50 2007 From: xiaw from umbi.umd.edu (wei xia) Date: Thu Nov 8 13:52:08 2007 Subject: [Zbrafish] Help with microinjected mRNA Message-ID: <38355AF6-DC46-4D94-BD31-6D0A0B773062@umbi.umd.edu> Hi Tony, Sorry for the bothering to other peoples. I have the same problem you had before. I injected in vitro transcribed capped GFP mRNA (Ambio mMessage Kit) into zebrafish 2-cell-stage embryos but I got no expression from it. Instead when I injected the plasmid templates which I used for the transcription I got strong fluorescence. I saw Bruce's reply and he said " 24hpf embryo should light up like a christmas tree." But why I couldn't! How could you solve your problems? Any suggestions are very very great appreciated! Wei Following is your post. Hi all, > > I am new to microinjection of zebrafish embryos and I need help. > > Briefly, I have injected 200pg of capped EGFP-coding RNA and the > embryos show no expression. In addition, their yolk kind of leaks out/ > burst between 4hpf and 10hpf, thus killing the embryo.While uninjected > ones look fine. > > To help with understanding the problem, I have detailed my protocol > here. > > Capped RNA Synthesis > > I linearized pSP64TNE-EGFP at the Sal I site, Phenyl-Chloroform > extracted the linearized template and then transcribe capped RNA with > SP6 promoter using the Ambion mMessege Kit. The yield comes in at an > acceptable 21ug with A260/280 at 1.94 and a A260/230 at 2.36. Running > 200ng of the capped RNA in a 1% agarose gel gives me a sharp single > band. > > Microinjection > > I diluted the capped RNA to 50ng/ul in 0.1M KCl and 0.05% PhenolRed. > 4nl(200pg) was injected into each embryo, the embryos are recovered > into 30% Danieau's Solution, and keep at 28.5C incubator. > > Any idea what can I be doing wrong here? > > Thanks a million for reading. > > Tony -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20071108/9cbb3c48/attachment.html From cheuk.chung from gmail.com Thu Nov 8 21:59:52 2007 From: cheuk.chung from gmail.com (Tony Cheuk-Chung Ho) Date: Fri Nov 9 12:14:34 2007 Subject: [Zbrafish] Re: Help with microinjected mRNA In-Reply-To: References: Message-ID: <1194577192.622492.172060@e9g2000prf.googlegroups.com> Wei, After reading Bruce's advise (refer to my previous post http://groups.google.com/group/bionet.organisms.zebrafish/browse_thread/thread/5c2bd01c334f425a), I precipitate my capped RNA and resuspend in RNase-free water. At this point, I tried to make them as concentrated as possible, maybe around 500ng/uL. Before injection, I mix the 500ng/uL stock with 0.05% Phenol Red in RNase-free water. Then, I inject embryos at 1-2 cell stage with 0.5, 1nL, and 2nL. This seems to be working fine for me as expression goes. Though not as "christmas tree" as I have wished, but expression is crisp and strong at 24hpf. I was able to observe weak fluorescence signal as early as 4-5hpf. Hope it helps. Tony On Nov 9, 2:46 am, wei xia wrote: > Hi Tony, > Sorry for the bothering to other peoples. I have the same problem you > had before. I injected in vitro transcribed capped GFP mRNA (Ambio > mMessage Kit) into zebrafish 2-cell-stage embryos but I got no > expression from it. Instead when I injected the plasmid templates > which I used for the transcription I got strong fluorescence. I saw > Bruce's reply and he said " 24hpf embryo should light up like a > christmas tree." But why I couldn't! > How could you solve your problems? > Any suggestions are very very great appreciated! > Wei > > Following is your post. > Hi all, > > > > I am new to microinjection of zebrafish embryos and I need help. > > > > Briefly, I have injected 200pg of capped EGFP-coding RNA and the > > embryos show no expression. In addition, their yolk kind of leaks > out/ > > burst between 4hpf and 10hpf, thus killing the embryo.While > uninjected > > ones look fine. > > > > To help with understanding the problem, I have detailed my protocol > > here. > > > > Capped RNA Synthesis > > > > I linearized pSP64TNE-EGFP at the Sal I site, Phenyl-Chloroform > > extracted the linearized template and then transcribe capped RNA with > > SP6 promoter using the Ambion mMessege Kit. The yield comes in at an > > acceptable 21ug with A260/280 at 1.94 and a A260/230 at 2.36. Running > > 200ng of the capped RNA in a 1% agarose gel gives me a sharp single > > band. > > > > Microinjection > > > > I diluted the capped RNA to 50ng/ul in 0.1M KCl and 0.05% PhenolRed. > > 4nl(200pg) was injected into each embryo, the embryos are recovered > > into 30% Danieau's Solution, and keep at 28.5C incubator. > > > > Any idea what can I be doing wrong here? > > > > Thanks a million for reading. > > > > Tony From wclements from ucsd.edu Fri Nov 9 13:18:05 2007 From: wclements from ucsd.edu (Wilson Clements) Date: Fri Nov 9 18:37:34 2007 Subject: [Zbrafish] Re: Zbrafish Digest, Vol 30, Issue 4 In-Reply-To: <200711091704.lA9H4MY17605@net.bio.net> References: <200711091704.lA9H4MY17605@net.bio.net> Message-ID: Dear Tony and Wei, There is a link to an excellent protocol with good detail (from the Mullins lab) at the following website. This protocol might help you troubleshoot your injections. http://www.courses.mbl.edu/zebrafish/faculty/mullins/ Personally, I don't dilute my mRNA into 0.1M KCl, but rather just inject it in straight DEPC water. Also, I use E3 media (from the zebrafish book, Westerfield et al.) to recover the injected embryos in. I have never tried raising embryos in Danieau's buffer, but it seems pretty high salt: could it be causing them to exogastrulate? Injection trauma is also hard on the embryos. How are you pulling your needles? Are they very small bore? What volume are you injecting? A 1nl drop should be maybe a 1/5 or 1/6 the volume of the single blastomere at the 1-cell stage. Large needles and large injection volumes will definitely kill the embryos (not to mention deliver way too much nucleic acid). Have you considered that it might not be an injection problem, but a scope problem? Are you using the right fluorescence filters? Is the lamp working? Do you have a positive control? Best, Wilson ------------------------------------------------------------------------ -------------------- Wilson Clements, Ph.D. wclements@ucsd.edu Dept. of Biology Section of Cell and Developmental Biology University of California at San Diego 9500 Gilman Dr. Natural Sciences Building 6105 La Jolla, CA 92093-0380 TEL (858) 534-6955 LAB (858) 822-4658 FAX (858) 822-5740 On Nov 9, 2007, at 9:04 AM, zbrafish-request@oat.bio.indiana.edu wrote: > Send Zbrafish mailing list submissions to > zbrafish@net.bio.net > > To subscribe or unsubscribe via the World Wide Web, visit > http://www.bio.net/biomail/listinfo/zbrafish > or, via email, send a message with subject or body 'help' to > zbrafish-request@net.bio.net > > You can reach the person managing the list at > zbrafish-owner@net.bio.net > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Zbrafish digest..." > > > Today's Topics: > > 1. Help Help with microinjecting RNA (wei xia) > 2. Help with microinjected mRNA (wei xia) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Thu, 8 Nov 2007 13:33:30 -0500 > From: wei xia > Subject: [Zbrafish] Help Help with microinjecting RNA > To: zbrafish@magpie.bio.indiana.edu > Message-ID: <4BEB7629-6442-4A04-A836-5AB1B8DB8985@umbi.umd.edu> > Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed > > Can you please forward my request to the author of this post? Cause I > have the same problem that he had met and I really can't figure it > out now. > Thank you very much for your help. > Wei > > > > ------------------------------ > > Message: 2 > Date: Thu, 8 Nov 2007 13:46:50 -0500 > From: wei xia > Subject: [Zbrafish] Help with microinjected mRNA > To: zbrafish@magpie.bio.indiana.edu > Message-ID: <38355AF6-DC46-4D94-BD31-6D0A0B773062@umbi.umd.edu> > Content-Type: text/plain; charset="us-ascii" > > Hi Tony, > Sorry for the bothering to other peoples. I have the same problem you > had before. I injected in vitro transcribed capped GFP mRNA (Ambio > mMessage Kit) into zebrafish 2-cell-stage embryos but I got no > expression from it. Instead when I injected the plasmid templates > which I used for the transcription I got strong fluorescence. I saw > Bruce's reply and he said " 24hpf embryo should light up like a > christmas tree." But why I couldn't! > How could you solve your problems? > Any suggestions are very very great appreciated! > Wei > > Following is your post. > Hi all, >> >> I am new to microinjection of zebrafish embryos and I need help. >> >> Briefly, I have injected 200pg of capped EGFP-coding RNA and the >> embryos show no expression. In addition, their yolk kind of leaks > out/ >> burst between 4hpf and 10hpf, thus killing the embryo.While > uninjected >> ones look fine. >> >> To help with understanding the problem, I have detailed my protocol >> here. >> >> Capped RNA Synthesis >> >> I linearized pSP64TNE-EGFP at the Sal I site, Phenyl-Chloroform >> extracted the linearized template and then transcribe capped RNA with >> SP6 promoter using the Ambion mMessege Kit. The yield comes in at an >> acceptable 21ug with A260/280 at 1.94 and a A260/230 at 2.36. Running >> 200ng of the capped RNA in a 1% agarose gel gives me a sharp single >> band. >> >> Microinjection >> >> I diluted the capped RNA to 50ng/ul in 0.1M KCl and 0.05% PhenolRed. >> 4nl(200pg) was injected into each embryo, the embryos are recovered >> into 30% Danieau's Solution, and keep at 28.5C incubator. >> >> Any idea what can I be doing wrong here? >> >> Thanks a million for reading. >> >> Tony > -------------- next part -------------- > An HTML attachment was scrubbed... > URL: http://www.bio.net/bionet/mm/zbrafish/attachments/ > 20071108/9cbb3c48/attachment-0001.html > > ------------------------------ > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > > End of Zbrafish Digest, Vol 30, Issue 4 > *************************************** > -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20071109/d6f5c288/attachment-0001.html From noopur.mandrekar from gmail.com Mon Nov 12 05:03:19 2007 From: noopur.mandrekar from gmail.com (Noopur) Date: Mon Nov 12 12:12:59 2007 Subject: [Zbrafish] Zebrafish liver toxicity Message-ID: <1194861799.536094.215860@e9g2000prf.googlegroups.com> Hi, I am Noopur Mandrekar from India. I am studying Zebrafish liver toxicity. Can anyone tell me the difference between Adult Zebrafish SGPT enzyme and human SGPT enzyme? Can we use this model to study liver toxicity using the biochemical parameters. Is there any other method to look out for Zebrafish liver toxicity in adult fish or embryonic larvae. Please let me know ASAP. Thanking you in advance. Best Regards, Noopur Mandrekar From wclements from ucsd.edu Fri Nov 9 13:18:05 2007 From: wclements from ucsd.edu (Wilson Clements) Date: Mon Nov 12 12:13:23 2007 Subject: [Zbrafish] Re: Zbrafish Digest, Vol 30, Issue 4 In-Reply-To: <200711091704.lA9H4MY17605@net.bio.net> References: <200711091704.lA9H4MY17605@net.bio.net> Message-ID: Dear Tony and Wei, There is a link to an excellent protocol with good detail (from the Mullins lab) at the following website. This protocol might help you troubleshoot your injections. http://www.courses.mbl.edu/zebrafish/faculty/mullins/ Personally, I don't dilute my mRNA into 0.1M KCl, but rather just inject it in straight DEPC water. Also, I use E3 media (from the zebrafish book, Westerfield et al.) to recover the injected embryos in. I have never tried raising embryos in Danieau's buffer, but it seems pretty high salt: could it be causing them to exogastrulate? Injection trauma is also hard on the embryos. How are you pulling your needles? Are they very small bore? What volume are you injecting? A 1nl drop should be maybe a 1/5 or 1/6 the volume of the single blastomere at the 1-cell stage. Large needles and large injection volumes will definitely kill the embryos (not to mention deliver way too much nucleic acid). Have you considered that it might not be an injection problem, but a scope problem? Are you using the right fluorescence filters? Is the lamp working? Do you have a positive control? Best, Wilson ------------------------------------------------------------------------ -------------------- Wilson Clements, Ph.D. wclements@ucsd.edu Dept. of Biology Section of Cell and Developmental Biology University of California at San Diego 9500 Gilman Dr. Natural Sciences Building 6105 La Jolla, CA 92093-0380 TEL (858) 534-6955 LAB (858) 822-4658 FAX (858) 822-5740 On Nov 9, 2007, at 9:04 AM, zbrafish-request@oat.bio.indiana.edu wrote: > Send Zbrafish mailing list submissions to > zbrafish@net.bio.net > > To subscribe or unsubscribe via the World Wide Web, visit > http://www.bio.net/biomail/listinfo/zbrafish > or, via email, send a message with subject or body 'help' to > zbrafish-request@net.bio.net > > You can reach the person managing the list at > zbrafish-owner@net.bio.net > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Zbrafish digest..." > > > Today's Topics: > > 1. Help Help with microinjecting RNA (wei xia) > 2. Help with microinjected mRNA (wei xia) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Thu, 8 Nov 2007 13:33:30 -0500 > From: wei xia > Subject: [Zbrafish] Help Help with microinjecting RNA > To: zbrafish@magpie.bio.indiana.edu > Message-ID: <4BEB7629-6442-4A04-A836-5AB1B8DB8985@umbi.umd.edu> > Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed > > Can you please forward my request to the author of this post? Cause I > have the same problem that he had met and I really can't figure it > out now. > Thank you very much for your help. > Wei > > > > ------------------------------ > > Message: 2 > Date: Thu, 8 Nov 2007 13:46:50 -0500 > From: wei xia > Subject: [Zbrafish] Help with microinjected mRNA > To: zbrafish@magpie.bio.indiana.edu > Message-ID: <38355AF6-DC46-4D94-BD31-6D0A0B773062@umbi.umd.edu> > Content-Type: text/plain; charset="us-ascii" > > Hi Tony, > Sorry for the bothering to other peoples. I have the same problem you > had before. I injected in vitro transcribed capped GFP mRNA (Ambio > mMessage Kit) into zebrafish 2-cell-stage embryos but I got no > expression from it. Instead when I injected the plasmid templates > which I used for the transcription I got strong fluorescence. I saw > Bruce's reply and he said " 24hpf embryo should light up like a > christmas tree." But why I couldn't! > How could you solve your problems? > Any suggestions are very very great appreciated! > Wei > > Following is your post. > Hi all, >> >> I am new to microinjection of zebrafish embryos and I need help. >> >> Briefly, I have injected 200pg of capped EGFP-coding RNA and the >> embryos show no expression. In addition, their yolk kind of leaks > out/ >> burst between 4hpf and 10hpf, thus killing the embryo.While > uninjected >> ones look fine. >> >> To help with understanding the problem, I have detailed my protocol >> here. >> >> Capped RNA Synthesis >> >> I linearized pSP64TNE-EGFP at the Sal I site, Phenyl-Chloroform >> extracted the linearized template and then transcribe capped RNA with >> SP6 promoter using the Ambion mMessege Kit. The yield comes in at an >> acceptable 21ug with A260/280 at 1.94 and a A260/230 at 2.36. Running >> 200ng of the capped RNA in a 1% agarose gel gives me a sharp single >> band. >> >> Microinjection >> >> I diluted the capped RNA to 50ng/ul in 0.1M KCl and 0.05% PhenolRed. >> 4nl(200pg) was injected into each embryo, the embryos are recovered >> into 30% Danieau's Solution, and keep at 28.5C incubator. >> >> Any idea what can I be doing wrong here? >> >> Thanks a million for reading. >> >> Tony > -------------- next part -------------- > An HTML attachment was scrubbed... > URL: http://www.bio.net/bionet/mm/zbrafish/attachments/ > 20071108/9cbb3c48/attachment-0001.html > > ------------------------------ > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > > End of Zbrafish Digest, Vol 30, Issue 4 > *************************************** > -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20071109/d6f5c288/attachment-0003.html From kinbaily from 126.com Tue Nov 13 16:30:37 2007 From: kinbaily from 126.com (kinbaily) Date: Tue Nov 13 18:00:54 2007 Subject: [Zbrafish] need adult zebrafish (6 months old) Message-ID: <20438464.10191194989437407.JavaMail.coremail@bj126app105.126.com> Hi, everyone, We need a lot of adult zebrafish (6 months old). We feel bad with the current provider and we have to change to other vendor. Any information about a reliable vendor will be highly appreciated. Thanks a lot. By the way, the price of Zebrafish International Resource Center (ZIRC) is too high for us. Thanks a lot. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20071114/5af17730/attachment.html From LStuemky from del-norte.k12.co.us Tue Nov 13 16:44:06 2007 From: LStuemky from del-norte.k12.co.us (Laura Stuemky) Date: Tue Nov 13 18:01:17 2007 Subject: [Zbrafish] zebrafish breeding Message-ID: <80C5BC73C024BF40964F3A814385F66CD87C29@DELNORTE8.del-norte.k12.co.us> I have a student wanting to do studies with zebrafish. His primary interest is cancer - but as we didn't have much luck finding a rat and mice lab for him to work in - he suggested this organism. DO you have any suggestions or advice for him? He is interested in exposing them to solutions from tobacco and cigarettes to see if there is a difference in the tumors produced. What type of time frame would this take to occur? Laura Stuemky Science teacher Del Norte High School Del Norte, CO -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20071113/730cda57/attachment.html From bdschyth from gmail.com Wed Nov 14 03:12:36 2007 From: bdschyth from gmail.com (bdschyth) Date: Wed Nov 14 13:23:07 2007 Subject: [Zbrafish] Re: zebrafish breeding In-Reply-To: References: Message-ID: <1195027956.143088.283230@50g2000hsm.googlegroups.com> On Nov 13, 10:44 pm, Laura Stuemky wrote: > I have a student wanting to do studies with zebrafish. His primary interest > is cancer - but as we didn't have much luck finding a rat and mice lab for > him to work in - he suggested this organism. DO you have any suggestions > or advice for him? He is interested in exposing them to solutions from > tobacco and cigarettes to see if there is a difference in the tumors > produced. What type of time frame would this take to occur? > Laura Stuemky > Science teacher > Del Norte High School > Del Norte, CO Dear Laura, My work is concerned with RNA interference and viral disease in rainbow trout so I do not have an exhaustive ref list on this but I do know that both rainbow trout and zebrafish have been used as cancer models. Recently I was at a phd defence regarding bladder cancer and we talked about the fish as model - as it was said by one - they dont smoke. Still Tobacco might have an effect on the fish as a model. Here is a 1996 review article retrieved from my ref manager about fish as models for environmental carcinogenesis: Bailey GS, Williams DE, Hendricks JD. 1996. Fish models for environmental carcinogenesis: the rainbow trout. Environ Health Perspect. Mar;104 Suppl 1:5-21. all the best Brian Dall Schyth, post. doc. Natl. Vet. Inst., Denmark From bdschyth from gmail.com Wed Nov 14 03:31:01 2007 From: bdschyth from gmail.com (bdschyth) Date: Wed Nov 14 13:23:29 2007 Subject: [Zbrafish] Re: RNAi in zebrafish In-Reply-To: References: Message-ID: <1195029061.855314.3400@d55g2000hsg.googlegroups.com> On Sep 18, 4:29 am, Joule wrote: > On Jul 24, 11:58 pm, senthil kumar wrote: > > > Is it possible to do RNAi in Zebrafish. Please send me the references.. > > Thanks > > Rsen > > > --------------------------------- > > Did you know? You can CHAT without downloading messenger. Click here > > It seems that RNAi does not work in zebrafish . ----------------------------------------------------------------------------------------------- But - why does it not work in fish? I dont think it is that well investigated yet. Most studies have been in zebrafish embryos microinjected with long dsRNAs. Remember that not many mammalian studies on RNAi mediated gene silencing has been publiched neither. Brian Dall Schyth, National Veterinary Institute, Denmark From jmoulton from gene-tools.com Wed Nov 14 13:44:27 2007 From: jmoulton from gene-tools.com (Jon D.Moulton) Date: Wed Nov 14 13:52:41 2007 Subject: [Zbrafish] Re: RNAi in zebrafish In-Reply-To: <1195029061.855314.3400@d55g2000hsg.googlegroups.com> References: <1195029061.855314.3400@d55g2000hsg.googlegroups.com> Message-ID: <473B420B.6040707@gene-tools.com> Regarding RNAi in zebrafish, this might be helpful: Gruber J, Manninga H, Tuschl T, Osborn M, Weber K. Specific RNAi mediated gene knockdown in zebrafish cell lines. RNA Biol. 2005 Jul-Sep;2(3):101-5. Epub 2005 Jul 19. The paper includes data on work in embryos, but they do not claim success in embryos with RNAi. Abstract: http://www.ncbi.nlm.nih.gov/sites/entrez?Db=pubmed&Cmd=ShowDetailView&TermToSearch=17114924 Open-access paper: http://www.landesbioscience.com/journals/rnabiology/article/gruberRNA2-3.pdf Let me know how I can help. Regards, - Jon Jon D. Moulton, Ph.D. Diagnostics and Special Projects GENE TOOLS, LLC jmoulton@gene-tools.com www.gene-tools.com (541) 929-7840 x1201 http://network.nature.com/group/morpholinos bdschyth wrote: > On Sep 18, 4:29 am, Joule wrote: > >> On Jul 24, 11:58 pm, senthil kumar wrote: >> >> >>> Is it possible to do RNAi in Zebrafish. Please send me the references.. >>> Thanks >>> Rsen >>> >>> --------------------------------- >>> Did you know? You can CHAT without downloading messenger. Click here >>> >> It seems that RNAi does not work in zebrafish . >> > ----------------------------------------------------------------------------------------------- > But - why does it not work in fish? I dont think it is that well > investigated yet. Most studies have been in zebrafish embryos > microinjected with long dsRNAs. Remember that not many mammalian > studies on RNAi mediated gene silencing has been publiched neither. > Brian Dall Schyth, National Veterinary Institute, Denmark > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20071114/c5f1638c/attachment.html From xiaw from umbi.umd.edu Fri Nov 16 15:32:56 2007 From: xiaw from umbi.umd.edu (xiawei) Date: Mon Nov 19 13:12:44 2007 Subject: [Zbrafish] Anybody has ever used IDT 27mer dicer substrate in zebrafish embryos. Message-ID: <0E0717BA-2AE1-4B55-8405-E604BB99ECE2@umbi.umd.edu> Just like the topic. Thanks Wei From thomas.bartman from cchmc.org Tue Nov 20 09:40:25 2007 From: thomas.bartman from cchmc.org (Thomas Bartman) Date: Tue Nov 20 12:54:44 2007 Subject: [Zbrafish] Mapcrosses and Positional Cloning Message-ID: <20071120144044.9D9F9E8047@mx3.chmccorp.cchmc.org> Hi all. We're having trouble with our positional cloning project, and we hoping for some input on the following two questions: 1) Are others noticing a great deal of misassembly to the genome still? It seems that every new release of the assembly, our markers jump all over. For example, on Zv5 we had two markers 500 kb apart, which went to 12 megabases apart on Zv6. We kept walking, and got down to 4 MB, but then Zv7 pushed those 7 MB apart. Furthermore, Zv7 has three of our favorite markers in order A-B-C on the genome, but our data has them A-C-B. (as an aside, we are looking at another gene, and have found it (all 800 bp) with the identical DNA sequence on two different chromosomes on Zv7. Even if these are redundant copies, how likely is it that they will match at 800 of 800 bp, or is this a sign of further misassembly?) 2) This is the crossing strategy we used. Are there any glaring errors in this that could be causing us problems or are we o.k.? a) A female AB carrying the mutation was crossed to a male WIK. Embryos were collected for positional cloning from this mapcross. However, we ran out of embryo DNA and the mapcross line got old and stopped laying well. So, b) A male from the above mapcross was backcrossed to a female WIK. We continue to collect embryos from this mapcross backcross. I wasn't sure if I read somewhere if it makes a great difference if the crosses to the mapping line are against females or males. Thanks for any advice, comments. Tom Thomas Bartman, M.D., Ph.D. Divisions of Neonatology, Pulmonary Biology, and Developmental Biology Cincinnati Children's Hospital Medical Center 3333 Burnet Ave, MLC 7009 Cincinnati, OH 45229-3039 Office: 513-636-9902 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20071120/9b5a59bd/attachment.html From bdschyth from gmail.com Wed Nov 21 06:58:27 2007 From: bdschyth from gmail.com (bdschyth) Date: Wed Nov 21 18:10:17 2007 Subject: [Zbrafish] Re: Anybody has ever used IDT 27mer dicer substrate in zebrafish embryos. References: Message-ID: <006807fa-562c-47e7-9238-d8a70e102380@b36g2000hsa.googlegroups.com> On Nov 16, 9:32 pm, xiawei wrote: > Just like the topic. > Thanks > Wei Dear Wei I don?t think anything has been published on this. Liu et al. 2005 in Develoment, growth and differentiation vol. 47, 323-331 have used in vitro Dicer cleaved RNAs as siRNAs without luck (nonspecific defects). I think they used a mammalian recombinant Dicer so it might be better to ad substrates to the embryos so that the fishs own dicer could perform its action in the cell. Brian From rburdine from Princeton.EDU Mon Nov 26 10:33:35 2007 From: rburdine from Princeton.EDU (Burdine, Rebecca D) Date: Mon Nov 26 12:50:52 2007 Subject: [Zbrafish] Mapcrosses and Positional Cloning In-Reply-To: <20071120144044.9D9F9E8047@mx3.chmccorp.cchmc.org> References: <20071120144044.9D9F9E8047@mx3.chmccorp.cchmc.org> Message-ID: <9A199EA7817D8844955074BB433A74A5409EED@MBCLUSTER.pu.win.princeton.edu> Hi Tom, 1) Yes yes and yes. Our regions get flipped over, flipped around, squeezed together, then spread onto different chromosomes. What we do is rely on the MGH, HS and GAT maps to set up our region, then turn to the Radiation hybrid maps to help flesh these out. Finally we see how well ensemble has assembled this region. If ensembl is off, we just blast and assemble our own contig that mimics what the genetic maps tell us is true. We also check synteny with other organisms which gives us more confidence in the Ensembl assembly or in our own. In the end we often get a region that seems reasonable and then hunt for candidates. At times, our mutant ended up being in the region assembled on Ensembl, but not where you would predict based on mapping. aside - at the last Zfish meeting, the people from the genome project said that if they get two identical stretches that assemble to different places, they leave both in until they can determine which is real. 2) I think your strategy is ok. We map Wik/AB and it has worked fine for us. The only things to keep an eye out for is that in some of our WIK mapping we see WIK contributing two different sizes for the Z markers. So we sometimes have three or four band patterns to score. This can get really complicated. Good luck, Becky --------------------------------------------------- Rebecca D. Burdine, Ph.D. Assistant Professor Dept. of Molecular Biology Princeton University Washington Road Mof 433 Princeton, NJ 08544 Phone: (609) 258-7515 Fax: (609) 258-1343 Email: rburdine@princeton.edu Admin Assistant: Cathy Falk (609) 258-1604 ________________________________ From: zbrafish-bounces@oat.bio.indiana.edu [mailto:zbrafish-bounces@oat.bio.indiana.edu] On Behalf Of Thomas Bartman Sent: Tuesday, November 20, 2007 9:40 AM To: Zbrafish@magpie.bio.indiana.edu Subject: [Zbrafish] Mapcrosses and Positional Cloning Hi all. We're having trouble with our positional cloning project, and we hoping for some input on the following two questions: 1) Are others noticing a great deal of misassembly to the genome still? It seems that every new release of the assembly, our markers jump all over. For example, on Zv5 we had two markers 500 kb apart, which went to 12 megabases apart on Zv6. We kept walking, and got down to 4 MB, but then Zv7 pushed those 7 MB apart. Furthermore, Zv7 has three of our favorite markers in order A-B-C on the genome, but our data has them A-C-B. (as an aside, we are looking at another gene, and have found it (all 800 bp) with the identical DNA sequence on two different chromosomes on Zv7. Even if these are redundant copies, how likely is it that they will match at 800 of 800 bp, or is this a sign of further misassembly?) 2) This is the crossing strategy we used. Are there any glaring errors in this that could be causing us problems or are we o.k.? a) A female AB carrying the mutation was crossed to a male WIK. Embryos were collected for positional cloning from this mapcross. However, we ran out of embryo DNA and the mapcross line got old and stopped laying well. So, b) A male from the above mapcross was backcrossed to a female WIK. We continue to collect embryos from this mapcross backcross. I wasn't sure if I read somewhere if it makes a great difference if the crosses to the mapping line are against females or males. Thanks for any advice, comments. Tom Thomas Bartman, M.D., Ph.D. Divisions of Neonatology, Pulmonary Biology, and Developmental Biology Cincinnati Children's Hospital Medical Center 3333 Burnet Ave, MLC 7009 Cincinnati, OH 45229-3039 Office: 513-636-9902 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20071126/2e1e1543/attachment.html From chi-bin.chien from neuro.utah.edu Mon Nov 26 13:12:25 2007 From: chi-bin.chien from neuro.utah.edu (Chi-Bin Chien) Date: Mon Nov 26 13:32:17 2007 Subject: [Zbrafish] Mapcrosses and Positional Cloning In-Reply-To: <9A199EA7817D8844955074BB433A74A5409EED@MBCLUSTER.pu.win.princeton.edu> References: <20071120144044.9D9F9E8047@mx3.chmccorp.cchmc.org> <9A199EA7817D8844955074BB433A74A5409EED@MBCLUSTER.pu.win.princeton.edu> Message-ID: Hi Tom, I'd only add a couple of things to Becky's response: -- It is worth contacting people at Sanger/Ensembl (there should be a feedback link on the webpage) to let them know the problems you see. Hopefully it will lead to a resolution in Zv8. -- For propagating the next generation of mapcross in cases where you have already rough-mapped, I suggest raising an incross from an existing map pair. This ensures that your allele system does not become any more complex, and with a high likelihood your current informative markers will also be informative in the next generation. (Since WIKs are not isogenic, outcrossing to WIK could introduce new marker alleles.) Chi-Bin Chien At 10:33 AM -0500 11/26/07, Burdine, Rebecca D wrote: >Content-class: urn:content-classes:message >Content-Type: multipart/alternative; > boundary="----_=_NextPart_001_01C83041.B5CF2406" > >Hi Tom, > >1) Yes yes and yes. Our regions get flipped over, flipped around, >squeezed together, then spread onto different chromosomes. What we >do is rely on the MGH, HS and GAT maps to set up our region, then >turn to the Radiation hybrid maps to help flesh these out. Finally >we see how well ensemble has assembled this region. If ensembl is >off, we just blast and assemble our own contig that mimics what the >genetic maps tell us is true. > >We also check synteny with other organisms which gives us more >confidence in the Ensembl assembly or in our own. > >In the end we often get a region that seems reasonable and then hunt >for candidates. At times, our mutant ended up being in the region >assembled on Ensembl, but not where you would predict based on >mapping. > >aside - at the last Zfish meeting, the people from the genome >project said that if they get two identical stretches that assemble >to different places, they leave both in until they can determine >which is real. > >2) I think your strategy is ok. We map Wik/AB and it has worked >fine for us. The only things to keep an eye out for is that in some >of our WIK mapping we see WIK contributing two different sizes for >the Z markers. So we sometimes have three or four band patterns to >score. This can get really complicated. > >Good luck, >Becky > >--------------------------------------------------- >Rebecca D. Burdine, Ph.D. >Assistant Professor >Dept. of Molecular Biology >Princeton University >Washington Road Mof 433 >Princeton, NJ 08544 > >Phone: (609) 258-7515 >Fax: (609) 258-1343 >Email: rburdine@princeton.edu >Admin Assistant: Cathy Falk (609) 258-1604 > > > >From: zbrafish-bounces@oat.bio.indiana.edu >[mailto:zbrafish-bounces@oat.bio.indiana.edu] On Behalf Of Thomas >Bartman >Sent: Tuesday, November 20, 2007 9:40 AM >To: Zbrafish@magpie.bio.indiana.edu >Subject: [Zbrafish] Mapcrosses and Positional Cloning > >Hi all. > >We're having trouble with our positional cloning project, and we >hoping for some input on the following two questions: > >1) Are others noticing a great deal of misassembly to the genome >still? It seems that every new release of the assembly, our markers >jump all over. For example, on Zv5 we had two markers 500 kb apart, >which went to 12 megabases apart on Zv6. We kept walking, and got >down to 4 MB, but then Zv7 pushed those 7 MB apart. Furthermore, >Zv7 has three of our favorite markers in order A-B-C on the genome, >but our data has them A-C-B. > >(as an aside, we are looking at another gene, and have found it (all >800 bp) with the identical DNA sequence on two different chromosomes >on Zv7. Even if these are redundant copies, how likely is it that >they will match at 800 of 800 bp, or is this a sign of further >misassembly?) > >2) This is the crossing strategy we used. Are there any glaring >errors in this that could be causing us problems or are we o.k.? > > a) A female AB carrying the mutation was crossed to a male >WIK. Embryos were collected for positional cloning from this >mapcross. However, we ran out of embryo DNA and the mapcross line >got old and stopped laying well. So, > > b) A male from the above mapcross was backcrossed to a female >WIK. We continue to collect embryos from this mapcross backcross. > >I wasn't sure if I read somewhere if it makes a great difference if >the crosses to the mapping line are against females or males. > >Thanks for any advice, comments. > >Tom > >Thomas Bartman, M.D., Ph.D. >Divisions of Neonatology, Pulmonary Biology, and Developmental Biology >Cincinnati Children's Hospital Medical Center >3333 Burnet Ave, MLC 7009 >Cincinnati, OH 45229-3039 > >Office: 513-636-9902 > > >_______________________________________________ >Zbrafish mailing list >Zbrafish@net.bio.net >http://www.bio.net/biomail/listinfo/zbrafish -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20071126/ff387bcd/attachment.html From alessaycy from hotmail.com Tue Nov 27 14:26:28 2007 From: alessaycy from hotmail.com (Alessa) Date: Tue Nov 27 14:30:59 2007 Subject: [Zbrafish] DAPT treatment? Message-ID: <7474d62c-d0eb-4be3-819e-91add724d536@s36g2000prg.googlegroups.com> Hi all, Did anybody try DAPT treatment in juvenile fish (couple weeks old)? DAPT is a Notch inhibitor. Thanks! From scalligaris from fcm.uncu.edu.ar Wed Nov 28 13:23:07 2007 From: scalligaris from fcm.uncu.edu.ar (=?iso-8859-1?Q?Sebasti=E1n_Calligaris?=) Date: Wed Nov 28 14:31:39 2007 Subject: [Zbrafish] NEW ZEBRA FISH FACILITIES Message-ID: <003e01c831eb$c4391510$0ea8698c@acer73b6f86142> Dear Tim Denning, My name is Sebasti?n D. Calligaris, I'm a posdoc scientist and I'm very interested in working with zebrafish. I'm looking for information about zebrafish laboratory disegn and facilities and I became happy when I found your message in Internet about new zebra fish faciliites from Thu May 23 10:00:49 EST 2002. Could you receive help? Could you start up with the zebra fish lab? Best regards, PhD Sebasti?n D. Calligaris Laboratorio de Biolog?a Celular y Molecular IHEM-CONICET Facultad de Ciencias M?dicas, Univ Nac de Cuyo Phone: (54) 261-4494143 Fax: (54) 261-4494117 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20071128/2a1e3bcb/attachment.html From david from zebrafish.org Wed Nov 28 15:03:11 2007 From: david from zebrafish.org (David Lains) Date: Wed Nov 28 16:45:29 2007 Subject: [Zbrafish] NEW ZEBRA FISH FACILITIES In-Reply-To: <003e01c831eb$c4391510$0ea8698c@acer73b6f86142> References: <003e01c831eb$c4391510$0ea8698c@acer73b6f86142> Message-ID: <03e101c831f9$b7494830$25dbd890$@org> Hello Sebastian Welcome to Zebrafish. I am very interested in this subject area and would be glad to help. Please feel free to contact me. Best Fishes David Lains <}}}>< Aquaculturist, Research Assistant Zebrafish International Resource Center 5274 University of Oregon Eugene, Or 97403 Email: david@zebrafish.org pH: (541) 346-6028 ext. 18 fax: (541) 346-6151 From: zbrafish-bounces@oat.bio.indiana.edu [mailto:zbrafish-bounces@oat.bio.indiana.edu] On Behalf Of Sebasti?n Calligaris Sent: Wednesday, November 28, 2007 10:23 AM To: zbrafish@magpie.bio.indiana.edu Subject: [Zbrafish] NEW ZEBRA FISH FACILITIES Dear Tim Denning, My name is Sebasti?n D. Calligaris, I'm a posdoc scientist and I'm very interested in working with zebrafish. I'm looking for information about zebrafish laboratory disegn and facilities and I became happy when I found your message in Internet about new zebra fish faciliites from Thu May 23 10:00:49 EST 2002. Could you receive help? Could you start up with the zebra fish lab? Best regards, PhD Sebasti?n D. Calligaris Laboratorio de Biolog?a Celular y Molecular IHEM-CONICET Facultad de Ciencias M?dicas, Univ Nac de Cuyo Phone: (54) 261-4494143 Fax: (54) 261-4494117 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20071128/28fdae02/attachment.html From koumound from upatras.gr Fri Nov 30 12:16:43 2007 From: koumound from upatras.gr (G. Koumoundouros) Date: Fri Nov 30 13:32:05 2007 Subject: [Zbrafish] Spawning problems Message-ID: <4750457B.3040301@upatras.gr> Hello to everybody, for the last 3 months we face significant problems with the spawning of our fish. This is a problem that occasionally appears to our facilities, without been possible to be explained. This time, due to a malfunction or the RO water hardness elevated to almost 30 odH and ph to 8,4. Could this be a reason why we face this problem? Has anyone noticed any periodicity in the zebrafish spawning? Any help is appreciated Thanks in advance Giorgos From clawrence from rics.bwh.harvard.edu Fri Nov 30 13:55:30 2007 From: clawrence from rics.bwh.harvard.edu (Christian Lawrence) Date: Fri Nov 30 14:00:47 2007 Subject: [Zbrafish] Spawning problems In-Reply-To: <4750457B.3040301@upatras.gr> References: <4750457B.3040301@upatras.gr> Message-ID: <84772d70711301055x5cd8ced0je2af9f79426bab40@mail.gmail.com> Giorgos, That is very hard, and it is probably a problem, especially if these levels were achieved rapidly. Zebrafish are loosely classified as a "hard water" species, but the values that I know they perform well under are closer to 10-13 dH (this is where we run them in our facility). 30 dH is very high and this is likely be the main problem. Running pH in the 8s is not a problem, unless of course you have ammonia in your system. At the end of the day, it is important to consider that there could be (and likely are) many factors underlying your spawning difficulties. Certainly a rapid swing to high hardness values could be the driver, but it may be something else that is more problematic, especially if you have had problems before your water controlling system malfunctioned. Zebrafish should not display a periodicity in spawning once they mature. If conditions and diet are right, they will only become senescent with old age or as a result of negative behavioral interactions. Feel free to email me off list and I am happy to go through things more thoroughly with you. Chris On 11/30/07, G. Koumoundouros wrote: > Hello to everybody, > for the last 3 months we face significant problems with the spawning of > our fish. This is a problem that occasionally appears to our facilities, > without been possible to be explained. > > This time, due to a malfunction or the RO water hardness elevated to > almost 30 odH and ph to 8,4. Could this be a reason why we face this > problem? > > Has anyone noticed any periodicity in the zebrafish spawning? > > Any help is appreciated > > Thanks in advance > > Giorgos > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > > The information transmitted in this electronic communication is intended only > for the person or entity to whom it is addressed and may contain confidential > and/or privileged material. Any review, retransmission, dissemination or other > use of or taking of any action in reliance upon this information by persons or > entities other than the intended recipient is prohibited. If you received this > information in error, please contact the Compliance HelpLine at 800-856-1983 and > properly dispose of this information. > > > -- Christian Lawrence Brigham and Women's Hospital Karp Family Research Laboratories, 06-004B One Blackfan Circle Boston, MA 02115 Tel: 617.355.9041 Fax: 617.355.9064 From claudia_hohn from hotmail.com Fri Nov 30 14:30:22 2007 From: claudia_hohn from hotmail.com (Claudia) Date: Fri Nov 30 14:33:05 2007 Subject: [Zbrafish] Re: NEW ZEBRA FISH FACILITIES References: Message-ID: <23c1b54f-51f7-438b-b97f-61bc66867b11@a39g2000pre.googlegroups.com> On Nov 28, 12:23 pm, Sebasti?n Calligaris wrote: > Dear Tim Denning, > > My name is Sebasti?n D. Calligaris, I'm a posdoc scientist and I'm very interested in working with zebrafish. I'm looking for information about zebrafish laboratory disegn and facilities and I became happy when I found your message in Internet about new zebra fish faciliites from Thu May 23 10:00:49 EST 2002. Could you receive help? Could you start up with the zebra fish lab? > Best regards, > > PhD Sebasti?n D. Calligaris > Laboratorio de Biolog?a Celular y Molecular > IHEM-CONICET > Facultad de Ciencias M?dicas, Univ Nac de Cuyo > Phone: (54) 261-4494143 > Fax: (54) 261-4494117 Hi Sebastian, I have some information how to start-up a zebrafish facility with little money. Also some easy protocols to follow for raising zebrafish. Let me know if you wish more information: claudia_hohn@hotmail.com