From haytcb from gmail.com Mon Feb 2 09:36:42 2009 From: haytcb from gmail.com (haytcb@gmail.com) Date: Mon Feb 2 12:21:29 2009 Subject: [Zbrafish] Re: Construct/transposase [ ] References: Message-ID: <33b5ae60-0a03-4106-ab39-4cb456b04041@v18g2000pro.googlegroups.com> well~This concentration should work well~~but I just wonder what you mean by cRNA... Actually, you should inject your constructs together with the transposase mRNA. Good luck :) On Jan 27, 10:56?pm, Jonathan Schumacher wrote: > Greetings, > > I am a rookie with zebrafish transgenics and have been given a project > to inject embryos with a construct in conjunction with transposase > cRNA. ?I don't see in the literature where the concentration of > construct & transposase are described. ?I've been using 20ng/uL for > both construct & transposase. ?Does this seem like a reasonable > concentration? ?I saw on another posting that Dr. Lister described > using ng/uL concntrations. > Thoughts? > > Many thanks! > > Jonathan Schumacher > Research and Development Scientist > Anatomic Pathology Molecular Diagnostics > ARUP Institute for Clinical & Experimental Pathology > 500 Chipeta Way > Salt Lake City, UT 84108 > Telephone: 801-583-2787 extension 2958 > Fax: 801-584-5109 > Email: schu...@aruplab.com From mwinandy from ulg.ac.be Mon Feb 2 03:51:50 2009 From: mwinandy from ulg.ac.be (Marie Winandy) Date: Mon Feb 2 12:21:51 2009 Subject: [Zbrafish] Re: making gels for embryo injections In-Reply-To: <200901301703.n0UH36806643@net.bio.net> References: <200901301703.n0UH36806643@net.bio.net> Message-ID: <200902020951500281.00233713@smtp.ulg.ac.be> Dear Jonathan, We don't use anymore those gels, but put the eggs next to a glass microscope slide, in a dry petri dish, taking away all the water. The eggs are maintained by the slide while you injcet them. After injection you just have to rinse with a wash bottle filled with egg water and collect the eggs in a new, clean petri dish. Good luck! M. Winandy Marie Winandy, PhD Université de Liège GIGA B34 - Zebrafish Platform Avenue de l'Hôpital 1 B-4000 Liège - Sart Tilman Belgique Tél: +32.4.366.99.71 +32.4.366.33.38 +32.476.97.25.33 Fax: +32.4.366.41.98 email : mwinandy@ulg.ac.be or zebrafish.giga@ulg.ac.be http://www.giga.ulg.ac.be/extranet/services.htm From tarini.sahoo from jyu.fi Sun Feb 1 08:43:05 2009 From: tarini.sahoo from jyu.fi (Tarini Prasad Sahoo) Date: Mon Feb 2 12:22:16 2009 Subject: [Zbrafish] Larval rearing Message-ID: <1932.130.234.109.128.1233495785.squirrel@webmail2.cc.jyu.fi> Hello everybody, I am new to the world of zf breeding as new can be! There was some success with getting the fish to spawn,but have had trouble with raising the fry. I followed the following method: -Day 0-6 the larvae were maintained in a 2L glass tray with no aeration. -Day 6,instead of the recommended 25-30 fry per 100 ml(250 ml beaker), I placed around 45 in 1L (2L beaker). No aeration here. Liquid diet (Nobil fluid artemia,JBL)as a substitute for infusoria diet. -On day 7, newly hatched artemia was given. No aeration here too,though the water was changed. -Day 8, 7 fry were found dead in the beaker, and the water was murky with foul odour. I moved the fry to a new beaker (600 ml in 2L) and minimal aeration through a glass pipette . 1.Would it be wise to resume the artemia diet only after regular aeration in the container/tank? 2.How many days post-fert can the fry withstand aeration? Since I have to use a static setup althrough,suggestions are welcome. Thank you in advance. Tarini Prasad Sahoo M.Sc,PhD student Department of Biological and Environmental Sciences Survontie 9 University of Jyv?skyl? Jyv?skyl?,FI-40014,Finland From elizabeth.laver from gmail.com Mon Feb 2 14:11:19 2009 From: elizabeth.laver from gmail.com (Elizabeth Laver) Date: Mon Feb 2 14:31:20 2009 Subject: [Zbrafish] four types of microinjection molds for $25 each Message-ID: <75615d510902021111h615a6444y3768f403fbc0199d@mail.gmail.com> We have purchased four different types of microinjection molds from Adaptive Science Tools at http://adaptivesciencetools.com/index.html and use them according to what we are doing with the embryos. All types of molds are $25.00 each. Zebrafish Injection: TU-1 = Six ramps containing one 90 degree and one 45 degree beveled side. Cnf-2 = Two ramps for holding dechorioned embryos used for time-lapse dorsal or ventral imaging Cell transplantation: PT-1= Injection/ transplantation mold with 150 triangular divots to accept individual embryos. I-34 = Six ramps used for micro-injection. Both sides are at a 90-degree angle firmly orienting the embryo. To order molds call (774) 239-6133 or email Inject@AdaptiveScienceTools.com They will also make molds and other bench top and fish facility equipment to your specifications. See the website for more details. Hope this helps. -Elizabeth On Fri, Jan 30, 2009 at 1:23 PM, Cecilia Moens wrote: > We purchased our microinjection molds from Clive Ellard Instrumentation in > Washington (360) 805-5406, clive@ellardinstrumentation.com. He sells a > mold for making agarose dishes with six parallel troughs for microinjection, > for $40.00 each. I'm not sure what the part number is, but tell them you > want the mold they made for the Moens lab at the Fred Hutchinson Cancer > Research Center. Also, Adaptive Science Tools in MA (774) 239-6133 sells a > plastic mold for making agarose dishes with 120 single wells for individual > embryos (part # PT-1). These are useful for transplantation experiments. > They may sell a mold for making troughs like the one from Clive Ellard, but > I'm not certain about this. > > - Cecilia. > > Cecilia B. Moens > > Division of Basic Science > > Fred Hutchinson Cancer Research Center > > B2-152, 1100 Fairview Ave. N. > > Seattle, WA 98109-1024 > > > office: (206) 667-5627 > > lab: (206) 667-5697 > > fax: (206) 667-3308 > > > _______________________________________________ > 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/20090202/b5a8bf0b/attachment.html From elizabeth.laver from gmail.com Mon Feb 2 14:26:11 2009 From: elizabeth.laver from gmail.com (Elizabeth Laver) Date: Mon Feb 2 14:32:06 2009 Subject: [Zbrafish] four types of microinjection molds for $25 a piece Message-ID: <75615d510902021126x7b678f3aud58dd02996b3f134@mail.gmail.com> We have purchased four different types of microinjection molds from Adaptive Science Tools at http://adaptivesciencetools.com/index.html and use them according to what we are doing with the embryos. All types of molds are $25.00 each. Zebrafish Injection: TU-1 = Six ramps containing one 90 degree and one 45 degree beveled side. Cnf-2 = Two ramps for holding dechorioned embryos used for time-lapse dorsal or ventral imaging Cell transplantation: PT-1= Injection/ transplantation mold with 150 triangular divots to accept individual embryos. I-34 = Six ramps used for micro-injection. Both sides are at a 90-degree angle firmly orienting the embryo. To order molds call (774) 239-6133 or email Inject@AdaptiveScienceTools.com They will also make molds and other bench top and fish facility equipment to your specifications. See the website for more details. Hope this helps. -Elizabeth On Fri, Jan 30, 2009 at 1:23 PM, Cecilia Moens wrote: > We purchased our microinjection molds from Clive Ellard Instrumentation in > Washington (360) 805-5406, clive@ellardinstrumentation.com. He sells a > mold for making agarose dishes with six parallel troughs for microinjection, > for $40.00 each. I'm not sure what the part number is, but tell them you > want the mold they made for the Moens lab at the Fred Hutchinson Cancer > Research Center. Also, Adaptive Science Tools in MA (774) 239-6133 sells a > plastic mold for making agarose dishes with 120 single wells for individual > embryos (part # PT-1). These are useful for transplantation experiments. > They may sell a mold for making troughs like the one from Clive Ellard, but > I'm not certain about this. > > - Cecilia. > > Cecilia B. Moens > > Division of Basic Science > > Fred Hutchinson Cancer Research Center > > B2-152, 1100 Fairview Ave. N. > > Seattle, WA 98109-1024 > > > office: (206) 667-5627 > > lab: (206) 667-5697 > > fax: (206) 667-3308 > > > _______________________________________________ > 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/20090202/220c4ef7/attachment-0001.html From anne_streit from web.de Tue Feb 3 10:40:28 2009 From: anne_streit from web.de (Anne Streit) Date: Tue Feb 3 12:22:56 2009 Subject: [Zbrafish] breeding problems in winter?? Message-ID: <1236143466@web.de> Hallo everybody! I?m working with zebrafish since octobre last year. We keep them in two large 50 litre tanks, male and females separated as good as differenciation was possible ;-) I use Tecniplast breeding boxes for pairing. The first pairings in octobre/november/december worked well. Unfortunately in the new year the fish have not layed eggs up to now, although I?ve tried several times to pair them. I also made long pauses to avoid stress for the females. The females do not have very round bellys :-( Does anyone know if there is a seasonal dependency? I pair them off-system, means at room temperature and set up pairs in the afternoon for the next morning. I would be very grateful for all advices concerning breeding! :-) Thanks in advance best regards Anne __________________________________________________________________________ Verschicken Sie SMS direkt vom Postfach aus - in alle deutschen und viele ausl?ndische Netze zum gleichen Preis! https://produkte.web.de/webde_sms/sms From Christian.Lawrence from childrens.harvard.edu Tue Feb 3 12:39:52 2009 From: Christian.Lawrence from childrens.harvard.edu (Lawrence, Christian) Date: Tue Feb 3 12:51:51 2009 Subject: [Zbrafish] breeding problems in winter?? In-Reply-To: <1236143466@web.de> References: <1236143466@web.de> Message-ID: <54F197250C79354BA5A2D4584D8D9FDA3FAF6B941D@CHEXCCRV1.CHBOSTON.ORG> There should be no seasonal dependency of reproduction in controlled conditions. If your females don't have "big bellies", they won't breed. You've got to get their bellies big again, so to speak - if you can. It is likely that they have become senescent because of poor diet and/or static social conditions. Feed them a mixture of a live diet and processed feeds and mix and match holding conditions as much as possible. Sex segregation is generally unnecessary. They will still spawn when set up in pairs and this will help keep them cycling if you don't regularly breed them. Long pauses between events are also not necessarily helpful. Although you will find it in various places in the "literature" that zebrafish females need a resting period of up to 2 weeks between spawning events, this is not the case if they are in top reproductive condition. Christian Lawrence Aquatic Resources Program Children's Hospital Boston 320 Longwood Avenue Boston, MA 02115 617.919.2738 (office) 617.730.0836 (fax) christian.lawrence@childrens.harvard.edu -----Original Message----- From: zbrafish-bounces@oat.bio.indiana.edu [mailto:zbrafish-bounces@oat.bio.indiana.edu] On Behalf Of Anne Streit Sent: Tuesday, February 03, 2009 10:40 AM To: bionet-organisms-zebrafish@moderators.isc.org Subject: [Zbrafish] breeding problems in winter?? Hallo everybody! I?m working with zebrafish since octobre last year. We keep them in two large 50 litre tanks, male and females separated as good as differenciation was possible ;-) I use Tecniplast breeding boxes for pairing. The first pairings in octobre/november/december worked well. Unfortunately in the new year the fish have not layed eggs up to now, although I?ve tried several times to pair them. I also made long pauses to avoid stress for the females. The females do not have very round bellys :-( Does anyone know if there is a seasonal dependency? I pair them off-system, means at room temperature and set up pairs in the afternoon for the next morning. I would be very grateful for all advices concerning breeding! :-) Thanks in advance best regards Anne __________________________________________________________________________ Verschicken Sie SMS direkt vom Postfach aus - in alle deutschen und viele ausl?ndische Netze zum gleichen Preis! https://produkte.web.de/webde_sms/sms _______________________________________________ Zbrafish mailing list Zbrafish@net.bio.net http://www.bio.net/biomail/listinfo/zbrafish The information in this e-mail is intended only for the person to whom it is addressed. If you believe this e-mail was sent to you in error and the e-mail contains patient information, please contact the Partners Compliance HelpLine at http://www.partners.org/complianceline . If the e-mail was sent to you in error but does not contain patient information, please contact the sender and properly dispose of the e-mail. From weibinz from med.umich.edu Wed Feb 4 16:38:20 2009 From: weibinz from med.umich.edu (Weibin Zhou) Date: Wed Feb 4 16:39:32 2009 Subject: [Zbrafish] BrdU for adult zebrafish Message-ID: <4989C480.8167.0008.0@med.umich.edu> Hi, I am wondering whether any one would be willing to share the protocol for BrdU labeling for adult zebrafish? and any recommendtion for BrdU antibody? Thank you. weibin ********************************************************** Electronic Mail is not secure, may not be read every day, and should not be used for urgent or sensitive issues From jeedward from yahoo.com Wed Feb 4 16:43:44 2009 From: jeedward from yahoo.com (John Edward) Date: Wed Feb 4 16:51:08 2009 Subject: [Zbrafish] Draft paper submission deadline extended: BCBGC-09 Message-ID: <221901.30840.qm@web45904.mail.sp1.yahoo.com> Draft paper submission deadline extended: BCBGC-09 ? The deadline for draft paper submission at the 2009 International Conference on Bioinformatics, Computational Biology, Genomics and Chemoinformatics (BCBGC-09) (website: http://www.PromoteResearch.org ) is extended due to numerous requests from the authors. The conference will be held during July 13-16 2009 in Orlando, FL, USA. We invite draft paper submissions. The conference will take place at the same time and venue where several other international conferences are taking place. The other conferences include: ????????? International Conference on Artificial Intelligence and Pattern Recognition (AIPR-09) ????????? International Conference on Automation, Robotics and Control Systems (ARCS-09) ????????? International Conference on Enterprise Information Systems and Web Technologies (EISWT-09) ????????? International Conference on High Performance Computing, Networking and Communication Systems (HPCNCS-09) ????????? International Conference on Information Security and Privacy (ISP-09) ????????? International Conference on Recent Advances in Information Technology and Applications (RAITA-09) ????????? International Conference on Software Engineering Theory and Practice (SETP-09) ????????? International Conference on Theory and Applications of Computational Science (TACS-09) ????????? International Conference on Theoretical and Mathematical Foundations of Computer Science (TMFCS-09) ? The website http://www.PromoteResearch.org contains more details. ? Sincerely John Edward Publicity committee ? -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090204/5d43e35f/attachment.html From mwinandy from ulg.ac.be Thu Feb 5 11:13:36 2009 From: mwinandy from ulg.ac.be (Marie Winandy) Date: Thu Feb 5 11:44:49 2009 Subject: [Zbrafish] zebrafish cell line In-Reply-To: <200902041703.n14H3J805663@net.bio.net> References: <200902041703.n14H3J805663@net.bio.net> Message-ID: <200902051713360781.01545E56@smtp.ulg.ac.be> Hello, I'm looking for someone who could share a zebrafish cell line and the tips for the culture ! I'm located in Belgium (Europe). This cell line should be easy to take care, it doesn't matter if it grows in layer or in suspension. As an alternative, can you tell me how to create one (I can obtain eggs or adult fish quite easily)? Thanking you in advance, Best regards, M. Winandy Marie Winandy, PhD Université de Liège GIGA B34 - Zebrafish Platform Avenue de l'Hôpital 1 B-4000 Liège - Sart Tilman Belgique Tél: +32.4.366.99.71 +32.4.366.33.38 +32.476.97.25.33 Fax: +32.4.366.41.98 email : mwinandy@ulg.ac.be or zebrafish.giga@ulg.ac.be http://www.giga.ulg.ac.be/extranet/services.htm From keviraj from yahoo.com Thu Feb 5 12:02:11 2009 From: keviraj from yahoo.com (raj abi) Date: Thu Feb 5 15:04:39 2009 Subject: [Zbrafish] Egg disinfection procedure Message-ID: <680153.65472.qm@web30408.mail.mud.yahoo.com> Hello, ? Could anyone pls suggest a method to?disinfect the zebra fish eggs?just to increase the hatching rate? ? best regards Rajesh Dr.Gerlai Lab University of Toronto -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090205/d08183c5/attachment.html From april from zebrafish.org Thu Feb 5 21:31:07 2009 From: april from zebrafish.org (April) Date: Fri Feb 6 12:39:50 2009 Subject: [Zbrafish] Egg disinfection procedure In-Reply-To: <680153.65472.qm@web30408.mail.mud.yahoo.com> References: <680153.65472.qm@web30408.mail.mud.yahoo.com> Message-ID: <3799B89D-7C9D-47E7-B79F-C515047DDBA5@zebrafish.org> Hi Rajesh, We have a recently updated our egg bleaching protocol which can be found on our website, http://zebrafish.org/zirc/documents/ protocols.php#Egg%20Bleaching. We have added a 1 minute pronase dip to the protocol which has increased our hatching rate to nearly 100%. If you have any questions about the protocol, please let me know. Very best, April On Feb 5, 2009, at 9:02 AM, raj abi wrote: > Hello, > > Could anyone pls suggest a method to disinfect the zebra fish eggs > just to increase the hatching rate? > > best regards > Rajesh > Dr.Gerlai Lab > University of Toronto > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish ----------------------- April R. Freeman (Mazanec) ZIRC Manager Zebrafish Int.'l Resource Center 5274 University of Oregon Eugene, OR 97403-5274 Phone: (541) 346-6028 ext. 24 Fax: (541) 346-6151 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090205/993bf80e/attachment.html From generalov_sergej from mail.ru Thu Feb 5 13:03:15 2009 From: generalov_sergej from mail.ru (Sergej Generalov) Date: Fri Feb 6 13:47:22 2009 Subject: [Zbrafish] RED Danio Message-ID: We have created a new genetically modified fish Danio rerio. It is an analog of GloFish Starfire Red Zebra. We use Fluorescent Proteins TurboFP635 to create a new fish. We have a line and start to produce a commercial quantity of the fish at the moment. Sergej From smitra from abdn.ac.uk Mon Feb 9 13:39:06 2009 From: smitra from abdn.ac.uk (Mitra, Suman) Date: Mon Feb 9 13:42:48 2009 Subject: [Zbrafish] Dual colour insitu protocol Message-ID: Hiya, Does anyone can help me with dual colour insitu protocol in ZF!! I will aprreciate any suggestion regarding this topic. Cheers, suman The University of Aberdeen is a charity registered in Scotland, No SC013683. From theresa.fresquez from state.nm.us Mon Feb 9 19:17:31 2009 From: theresa.fresquez from state.nm.us (Fresquez, Theresa, DCA) Date: Tue Feb 10 12:18:33 2009 Subject: [Zbrafish] (no subject) Message-ID: <1ACBC54813C0A34195711085D2DE3CF0072CA8BD@CEXMB2.nmes.lcl> Hello, Does anyone know how to get rid of coleps? They are eating my zebrafish babies. Thanks, Tess Confidentiality Notice: This e-mail, including all attachments is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited unless specifically provided under the New Mexico Inspection of Public Records Act. If you are not the intended recipient, please contact the sender and destroy all copies of this message. -- This email has been scanned by the Sybari - Antigen Email System. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090209/1067f0f4/attachment.html From april from zebrafish.org Tue Feb 10 15:15:14 2009 From: april from zebrafish.org (April) Date: Tue Feb 10 15:17:25 2009 Subject: [Zbrafish] (no subject) In-Reply-To: <1ACBC54813C0A34195711085D2DE3CF0072CA8BD@CEXMB2.nmes.lcl> References: <1ACBC54813C0A34195711085D2DE3CF0072CA8BD@CEXMB2.nmes.lcl> Message-ID: <26F689E7-4258-4CC5-88DB-8E741FE2D76A@zebrafish.org> Hi Tess, Here is my recommendation for containing Coleps and/or any other unwanted organisms. 1. Bleach all of your eggs as soon as possible. If needed, you can reference the egg bleaching protocol found in The Zebrafish Book, http://zfin.org/zf_info/zfbook/chapt3/3.2.html or on our website, http://zebrafish.org/zirc/documents/protocols.php#Egg%20Bleaching. 2. Be sure to remove ALL dead/damaged eggs before bleaching. 3. After bleaching, keep your embryos in embryo media until they have inflated their swim bladders and are swimming. Zebrafish larvae are the most susceptible when they have hatched, but not yet inflated their swim bladders. Once the larvae are able to swim properly, they can fend off most organisms. If you haven't already positively identified the organisms and would like to, I recommend putting a few of these critters on a slide and look at them under a compound microscope. Coleps are quite small and move in an intense twirling motion. They have plates that cover their exterior and make them look like hand grenades. As you mentioned, they are voracious and can consume an entire embryo in 45 minutes. For future reference, I recommend obtaining a good protozoa picture book/guide. We have 2 older books that are quite helpful, however you might be able to find some newer better books with even better pictures and information. - Patterson, D.J. (1996). Free-Living Freshwater Protozoa (A Colour Guide). John Wiley & Sons Inc., New York, New York. - Berk, S. G.; Gunderson, J. H (1993). Wastewater Organisms (A Color Atlas). CRC Press LLC. If you continue to have problems, please let me know. Very best, April On Feb 9, 2009, at 4:17 PM, Fresquez, Theresa, DCA wrote: > Hello, > Does anyone know how to get rid of coleps? They are eating my > zebrafish babies. > Thanks, > Tess > > > > > > Confidentiality Notice: This e-mail, including all attachments is > for the sole use of the intended recipient(s) and may contain > confidential and privileged information. Any unauthorized review, > use, disclosure or distribution is prohibited unless specifically > provided under the New Mexico Inspection of Public Records Act. If > you are not the intended recipient, please contact the sender and > destroy all copies of this message. -- This email has been scanned > by the Sybari - Antigen Email System. > > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish ----------------------- April R. Freeman (Mazanec) ZIRC Manager Zebrafish Int.'l Resource Center 5274 University of Oregon Eugene, OR 97403-5274 Phone: (541) 346-6028 ext. 24 Fax: (541) 346-6151 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090210/4b04e1fa/attachment.html From smitra from abdn.ac.uk Wed Feb 11 06:05:44 2009 From: smitra from abdn.ac.uk (Mitra, Suman) Date: Wed Feb 11 12:30:12 2009 Subject: [Zbrafish] Need insitu probes Message-ID: Hiya! Can any one share insitu probes for gata3, ikaros, lck, and the recombination activating genes rag-1 and rag-2 or provide me some guidence who all ready made this constructs. Cheers, suman The University of Aberdeen is a charity registered in Scotland, No SC013683. From rburdine from Princeton.EDU Wed Feb 11 12:38:26 2009 From: rburdine from Princeton.EDU (Burdine, Rebecca D) Date: Wed Feb 11 12:45:17 2009 Subject: [Zbrafish] Need insitu probes In-Reply-To: References: Message-ID: Hi Suman, I think the best and most appropriate approach is two pronged. 1) Check Zfin (www.zfin.org) to see if the probes are available for purchase. 2) Find the paper where the probe was originally used or determine from papers a lab that uses the probes regularly. The write to the corresponding author and ask for the probe from that lab directly. Some plasmids are given out under MTA agreements and may not be available for distribution from just anywhere. 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) 258-6730 Email: rburdine@princeton.edu Admin Assistant: Anna Schmedel (609) 258-5028 > -----Original Message----- > From: zbrafish-bounces@oat.bio.indiana.edu [mailto:zbrafish- > bounces@oat.bio.indiana.edu] On Behalf Of Mitra, Suman > Sent: Wednesday, February 11, 2009 6:06 AM > To: 'zbrafish@net.bio.net' > Subject: [Zbrafish] Need insitu probes > Importance: High > > Hiya! > Can any one share insitu probes for gata3, ikaros, lck, and the > recombination activating genes rag-1 and rag-2 or provide me some > guidence who all ready made this constructs. > > Cheers, > suman > > > The University of Aberdeen is a charity registered in Scotland, No > SC013683. > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish From mena22787 from excite.com Thu Feb 12 11:12:23 2009 From: mena22787 from excite.com (Serena) Date: Thu Feb 12 13:23:53 2009 Subject: [Zbrafish] question about peroxide block IHC/ISH Message-ID: <0f739ea0-215e-47ec-8acd-d0e78f17e79d@t13g2000yqc.googlegroups.com> I have a question about hte need for a peroxide block step in ISH. I have seen many whole mout protocols for IHC that require a peroxide block step to block endogenous peroxidases, however, the whole mount in situ protocols that I have seen do not have one, despite the similar AP detection chemistry. Can someone explain the reason for this? I am performing in situ on rather old embryos 10-15 days (for sectioning) and want to make sure I do not get a lot of background. Also, if anyone has experience with this technique on this age range, could you suggest a length of time for the proteinase K step. Thanks! From trevarrow from EugeneResearchAquatics.com Wed Feb 11 21:19:41 2009 From: trevarrow from EugeneResearchAquatics.com (Bill Trevarrow) Date: Thu Feb 12 13:24:13 2009 Subject: [Zbrafish] Re: Coleps In-Reply-To: <26F689E7-4258-4CC5-88DB-8E741FE2D76A@zebrafish.org> References: <1ACBC54813C0A34195711085D2DE3CF0072CA8BD@CEXMB2.nmes.lcl> <26F689E7-4258-4CC5-88DB-8E741FE2D76A@zebrafish.org> Message-ID: Hi Tess, I would add the following to the advice April provided: There are two common sources of Coleps predating upon fish eggs: 1) the water system or aquarium water the fish laying the eggs come from 2) the cultures of food organisms used to feed the baby fish (often paramecium) Introducing Coleps from either of these sources into your larval culturing system makes it possible for the Coleps to rapidly bloom if they are able to feed on an egg. If one of these is contaminated, it is pretty easy for the other to get contaminated with Coleps. Once your water system is contaminated, it will be very difficult to remove the Coleps from the water system without nuking the whole thing with bleach. Paramecium cultures can be restarted relatively easily. Normally, Coleps are saprophages, eating detritus. I guess fish eggs are just too enticing an object for them to pass up on. A dirty rather than a clean water system would therefore be expected to provide a better home for Coleps. This also provides another argument against overfeeding. Most commercially available cultures of ciliates are full of a variety of contaminants which should be removed such as: Coleps, Didinium (a predator on paramecia), and other fast growing organisms that (under some culture conditions) can out compete paramecia. The stock center however is a source of Coleps free paramecia cultures. Coleps and other unwanted organisms can be kept out of both sources by good quarantine procedures. This also means that introducing new feed organisms (such as new paramecium cultures) into your facility can also introduce Coleps. Since they can be seen in a dissecting microscope, it is possible to produce a Coleps free culture. A simple approach is: 1) dilute your initial culture 2) pick out individual organisms and put them into a new dish of culture media (diluting anything that came with them) 3) then pick out individuals again and put them into a new dish of culture media. You can start a culture with a single paramecium. Using more results in a large culture sooner, but is more work and is more likely to by chance include a contaminant. This process can be repeated several times, but two or three are usually sufficient to ensure the Coleps are not included. The more animals you transfer, the more work will be required, the greater possibility of a contaminant getting through, but also the more animals you have to start a new culture. I usually start two cultures in this way for a greater possibility of success. I also have written up another approach has been described in an old issue of the Zebrafish Monitor which may be available on the ZFIN.org website. Some claim a salt concentration shock can kill Coleps. Others say it is just a temporary effect that does not kill them all and that eventually they come back. The sudden salt change could also stress your fish. On Feb 10, 2009, at 12:15 PM, April wrote: > Hi Tess, > Here is my recommendation for containing Coleps and/or any other > unwanted organisms. > > 1. Bleach all of your eggs as soon as possible. If needed, you can > reference the egg bleaching protocol found in The Zebrafish Book, http://zfin.org/zf_info/zfbook/chapt3/3.2.html > or on our website, http://zebrafish.org/zirc/documents/protocols.php#Egg%20Bleaching > . > 2. Be sure to remove ALL dead/damaged eggs before bleaching. > 3. After bleaching, keep your embryos in embryo media until they > have inflated their swim bladders and are swimming. Zebrafish > larvae are the most susceptible when they have hnatched, but not yet > inflated their swim bladders. Once the larvae are able to swim > properly, they can fend off most organisms. > > If you haven't already positively identified the organisms and would > like to, I recommend putting a few of these critters on a slide and > look at them under a compound microscope. Coleps are quite small > and move in an intense twirling motion. They have plates that cover > their exterior and make them look like hand grenades. As you > mentioned, they are voracious and can consume an entire embryo in 45 > minutes. > > For future reference, I recommend obtaining a good protozoa picture > book/guide. We have 2 older books that are quite helpful, however > you might be able to find some newer better books with even better > pictures and information. > > - Patterson, D.J. (1996). Free-Living Freshwater Protozoa (A Colour > Guide). John Wiley & Sons Inc., New York, New York. > - Berk, S. G.; Gunderson, J. H (1993). Wastewater Organisms (A Color > Atlas). CRC Press LLC. > > If you continue to have problems, please let me know. > > Very best, > > April > > On Feb 9, 2009, at 4:17 PM, Fresquez, Theresa, DCA wrote: > >> Hello, >> Does anyone know how to get rid of coleps? They are eating my >> zebrafish babies. >> Thanks, >> Tess >> >> >> >> >> >> Confidentiality Notice: This e-mail, including all attachments is >> for the sole use of the intended recipient(s) and may contain >> confidential and privileged information. Any unauthorized review, >> use, disclosure or distribution is prohibited unless specifically >> provided under the New Mexico Inspection of Public Records Act. If >> you are not the intended recipient, please contact the sender and >> destroy all copies of this message. -- This email has been scanned >> by the Sybari - Antigen Email System. >> >> >> _______________________________________________ >> Zbrafish mailing list >> Zbrafish@net.bio.net >> http://www.bio.net/biomail/listinfo/zbrafish > > ----------------------- > April R. Freeman (Mazanec) > ZIRC Manager > Zebrafish Int.'l Resource Center > 5274 University of Oregon > Eugene, OR 97403-5274 > > Phone: (541) 346-6028 ext. 24 > Fax: (541) 346-6151 > > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish Bill Trevarrow, Ph.D. Eugene Research Aquatics, LLC trevarrow@EugeneResearchAquatics.com Tel: (541) 844-9054 Fax: (541) 683-0996 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090211/1f5a2cb4/attachment.html From rburdine from Princeton.EDU Thu Feb 12 14:22:19 2009 From: rburdine from Princeton.EDU (Burdine, Rebecca D) Date: Thu Feb 12 14:57:00 2009 Subject: [Zbrafish] question about peroxide block IHC/ISH In-Reply-To: <0f739ea0-215e-47ec-8acd-d0e78f17e79d@t13g2000yqc.googlegroups.com> References: <0f739ea0-215e-47ec-8acd-d0e78f17e79d@t13g2000yqc.googlegroups.com> Message-ID: <790DAC09623D0A47ACDB126E3C52D8B6019A64@MBCLUSTER.pu.win.princeton.edu> Hi Serena, Whether you need to block for endogenous AP really depends on the probe and the age of the embryos IMHO. For some weak or finicky probes, we find blocking endogenous AP is key. For really strong probes, there is typically no need since the signal over background is tremendously strong. For later embryos and larvae, I think it is important because the liver will light up like gangbusters without this step. In fact it will still light up with this step, but it is somewhat less. Unfortunately we have had no luck getting nice specific signals with whole-mount insitus on embryos older than 7 days, but this might be our probes. We tried sectioning after the protocol and that didn't work at all. I'd be interested in hearing if other labs have had good luck on whole-mount ISH in larvae that are this old. 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) 258-6730 Email: rburdine@princeton.edu Admin Assistant: Anna Schmedel (609) 258-5028 > -----Original Message----- > From: zbrafish-bounces@oat.bio.indiana.edu [mailto:zbrafish- > bounces@oat.bio.indiana.edu] On Behalf Of Serena > Sent: Thursday, February 12, 2009 11:12 AM > To: bionet-organisms-zebrafish@moderators.isc.org > Subject: [Zbrafish] question about peroxide block IHC/ISH > > I have a question about hte need for a peroxide block step in ISH. I > have seen many whole mout protocols for IHC that require a peroxide > block step to block endogenous peroxidases, however, the whole mount > in situ protocols that I have seen do not have one, despite the > similar AP detection chemistry. Can someone explain the reason for > this? I am performing in situ on rather old embryos 10-15 days (for > sectioning) and want to make sure I do not get a lot of background. > > Also, if anyone has experience with this technique on this age range, > could you suggest a length of time for the proteinase K step. > > Thanks! > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish From milka.sarris from gmail.com Fri Feb 13 07:29:12 2009 From: milka.sarris from gmail.com (MILKA) Date: Fri Feb 13 12:34:15 2009 Subject: [Zbrafish] number of cells of ZF 3-5d old embryos Message-ID: <5b53f651-75de-4be3-90b3-51f5cc645a1a@r41g2000yqm.googlegroups.com> Hello, Would anyone have an idea of how many total cells can be extracted from 3,4 or 5 d old ZF embryos with trypsin/EDTA-mediated dissociation? I am trying to prepare cell suspensions from ZF embryos and would like to have an idea of how many cells/embryo are possible to extract at the different developmental stages. Any insight would be very much appreciated! Many thanks, Milka From jimdowlingmdphd from mac.com Sat Feb 14 14:06:33 2009 From: jimdowlingmdphd from mac.com (Jim D.) Date: Mon Feb 16 12:11:32 2009 Subject: [Zbrafish] embryo/larval survival Message-ID: <8f3a442a-bcbc-43fc-95aa-3b3c13f5d862@i20g2000prf.googlegroups.com> Hello. I am planning a high throughput chemical screen on embryos/ larvae. I'd like to do either a 96 well or 24 well format. My question: does any one have a feel for how long an embryo can survive after dechorionation in a 96 or 24 well plate? If people have tried this, have they modified the plates at all (for example, poking holes in the lid). Thanks! Jim Dowling Assistant Professor, Department of Pediatrics and Neurology University of Michigan Medical Center From gilbert.weidinger from biotec.tu-dresden.de Mon Feb 16 18:56:19 2009 From: gilbert.weidinger from biotec.tu-dresden.de (Gilbert Weidinger) Date: Mon Feb 16 18:58:19 2009 Subject: [Zbrafish] AmCyan Message-ID: <4999FD23.3000408@biotec.tu-dresden.de> Hi all, Does anybody have experience with AmCyan in zebrafish? It's a cyan fluorescent protein from reef coral, part of the living color's series of Clonetech. I wonder how bright it is (relative to EGFP) and how fast it folds. I don't care that it is a tetramer, unless it forms large aggregates and thus might be toxic. Cerulean works well for us, but I am looking for something outside the GFP family since I'd like to be able to distinguish several FPs by antibodies. many thanks gilbert -- -------------------------------------------------------- Gilbert Weidinger, PhD Group Leader SFB 655 Biotechnology Center & Center for Regenerative Therapies University of Technology Dresden Mail address: Gilbert Weidinger Biotechnology Center Tatzberg 47-51 01307 Dresden Germany Tel. office: 0049 351 463 40120 Tel. lab: 0049 351 463 40108 Tel. secretary: 0049 351 463 40345 Fax: 0049 351 463 40348 email: gilbert.weidinger@biotec.tu-dresden.de web: http://www.biotec.tu-dresden.de/weidinger From fiona.lzht from gmail.com Mon Feb 16 21:17:14 2009 From: fiona.lzht from gmail.com (fiona.lzht@gmail.com) Date: Tue Feb 17 12:13:12 2009 Subject: [Zbrafish] everything is possible. Message-ID: Liu Zhaoting ,Ph.D. Dept. of State Key Laboratory of Biomembranes and Membrane Biotechnology Chinese Academy of Science Institute of Zoology Datun Road Beijing From padnos from yahoo.com Tue Feb 17 08:41:25 2009 From: padnos from yahoo.com (padnos@yahoo.com) Date: Tue Feb 17 12:13:29 2009 Subject: [Zbrafish] Re: embryo/larval survival References: Message-ID: <93ab964c-b657-4f70-bc26-10123b059796@i38g2000yqd.googlegroups.com> On Feb 14, 2:06?pm, "Jim D." wrote: > Hello. ?I am planning a high throughput chemical screen on embryos/ > larvae. ?I'd like to do either a 96 well or 24 well format. ?My > question: ?does any one have a feel for how long an embryo can survive > after dechorionation in a 96 or 24 well plate? ?If people have tried > this, have they modified the plates at all (for example, poking holes > in the lid). > Thanks! > > Jim Dowling > Assistant Professor, Department of Pediatrics and Neurology > University of Michigan Medical Center I raise fry daily from about 6 hpf to 6 dpf in 96 well plates (Millipore 96 well mesh plates which are now a special order) 1 per well and have about 95 to 100 % survival.periodically have riased in 24 and 48 well plates without any trouble to the same age with the same approximate result. We do not dechorinate. Beth Padnos US EPA RTP North Carolina From thebrog from hotmail.com Tue Feb 17 15:32:46 2009 From: thebrog from hotmail.com (thebrog@hotmail.com) Date: Tue Feb 17 15:37:39 2009 Subject: [Zbrafish] Any way to treat small lesions on fish? Message-ID: <7da11f08-36ec-4111-a21d-b8d71c36be38@j35g2000yqh.googlegroups.com> Just trying to find out any infomation on treating fish with small lesions that is effective and relatively quick. Any infomation would be great help. Thanks Andy From thebrog from hotmail.com Wed Feb 18 04:41:49 2009 From: thebrog from hotmail.com (thebrog@hotmail.com) Date: Wed Feb 18 12:34:37 2009 Subject: [Zbrafish] Any help on treating small leisons on zebrafish Message-ID: <94b789d9-0643-4fa6-b188-210acc94c71b@j38g2000yqa.googlegroups.com> Does anyone have any infomation or tips on treating zebrafish with small lesions? any info would be greatly received. thanks Andy From jeedward from yahoo.com Fri Feb 20 10:22:46 2009 From: jeedward from yahoo.com (John Edward) Date: Fri Feb 20 12:57:11 2009 Subject: [Zbrafish] Draft paper submission deadline extended: BCBGC-09 Message-ID: <134664.9191.qm@web45903.mail.sp1.yahoo.com> Draft paper submission deadline extended: BCBGC-09 ? The deadline for draft paper submission at the 2009 International Conference on Bioinformatics, Computational Biology, Genomics and Chemoinformatics (BCBGC-09) (website: http://www.PromoteResearch.org ) is extended due to numerous requests from the authors. The conference will be held during July 13-16 2009 in Orlando, FL, USA. We invite draft paper submissions. The conference will take place at the same time and venue where several other international conferences are taking place. The other conferences include: ????????? International Conference on Artificial Intelligence and Pattern Recognition (AIPR-09) ????????? International Conference on Automation, Robotics and Control Systems (ARCS-09) ????????? International Conference on Enterprise Information Systems and Web Technologies (EISWT-09) ????????? International Conference on High Performance Computing, Networking and Communication Systems (HPCNCS-09) ????????? International Conference on Information Security and Privacy (ISP-09) ????????? International Conference on Recent Advances in Information Technology and Applications (RAITA-09) ????????? International Conference on Software Engineering Theory and Practice (SETP-09) ????????? International Conference on Theory and Applications of Computational Science (TACS-09) ????????? International Conference on Theoretical and Mathematical Foundations of Computer Science (TMFCS-09) ? The website http://www.PromoteResearch.org contains more details. ? Sincerely John Edward Publicity committee -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090220/5ce4d5f8/attachment.html From l.bugeon from imperial.ac.uk Fri Feb 20 06:49:08 2009 From: l.bugeon from imperial.ac.uk (Bugeon, Laurence) Date: Fri Feb 20 12:57:37 2009 Subject: [Zbrafish] detergents Message-ID: <5C8D06A264FD1E4B8F88A7C31E5733F533D65CEC@ICEXM4.ic.ac.uk> Dear all, what are your favorite type of detergents to clean fish room floors and fish tanks? thanks for any advice. Laurence _______________________________________________ Zbrafish mailing list Zbrafish@net.bio.net http://www.bio.net/biomail/listinfo/zbrafish From rburdine from Princeton.EDU Fri Feb 20 15:17:24 2009 From: rburdine from Princeton.EDU (Burdine, Rebecca D) Date: Fri Feb 20 15:19:55 2009 Subject: [Zbrafish] detergents In-Reply-To: <5C8D06A264FD1E4B8F88A7C31E5733F533D65CEC@ICEXM4.ic.ac.uk> References: <5C8D06A264FD1E4B8F88A7C31E5733F533D65CEC@ICEXM4.ic.ac.uk> Message-ID: <790DAC09623D0A47ACDB126E3C52D8B6019D8D@MBCLUSTER.pu.win.princeton.edu> I'd love to hear answers to this question so I hope you will share any off-line responses. For tanks - we use nothing but RO water and elbow grease period. If tank really needs disinfecting, we do a dilute bleach rinse, RO rinse, and complete air drying. For the floors we use Simple Green. My former aquaculture technicians assure me this is the detergent used on the deep sea ships they interned on to clean the decks. It is relatively safe for aquatic life. (I wouldn't grow my fish in it for example, but for the floors it works fine.) 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) 258-6730 Email: rburdine@princeton.edu Admin Assistant: Anna Schmedel (609) 258-5028 > -----Original Message----- > From: zbrafish-bounces@oat.bio.indiana.edu [mailto:zbrafish- > bounces@oat.bio.indiana.edu] On Behalf Of Bugeon, Laurence > Sent: Friday, February 20, 2009 6:49 AM > To: bionet-organisms-zebrafish@moderators.isc.org > Subject: [Zbrafish] detergents > > > Dear all, > what are your favorite type of detergents to clean fish room floors and > fish tanks? > thanks for any advice. > Laurence > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish From gottalovelife from gmail.com Mon Feb 23 14:01:59 2009 From: gottalovelife from gmail.com (Kim) Date: Mon Feb 23 14:03:23 2009 Subject: [Zbrafish] Preparation of zebrafish embryos for oxidative stress assays Message-ID: <9e2446f8-88d8-47f2-b9d8-2f2339571289@v39g2000yqm.googlegroups.com> Hello all; Does anyone have experience preparing extract from zebrafish embryos for oxidative stress assays? From jaschumac from gmail.com Thu Feb 26 14:10:10 2009 From: jaschumac from gmail.com (Jonathan Schumacher) Date: Thu Feb 26 14:13:13 2009 Subject: [Zbrafish] Zfish injection/integration control Message-ID: Greetings, I have designed a construct of MITFA driving my oncogene and cytoplasmic-expressed EGFP, subsequently cloned into the pDest Tol2pA. We suspect that it will take at least 8 weeks to see whether the fish develop cancer and after setting up injections, I realized that I have no injection/integration control. My question is would you suggest recloning my construct into the pDest Tol2pA with cmlc2- driven EGFP? I believe that this would lead to double-EGFP labeling and am unclear if this would be problematic. Is there an alternative strategy, like coinjecting my destination vector with a cherry-labeled cmlc2 (or other)? Your thoughts & suggestions are greatly appreciated. Best, Jonathan From mingsanma from gmail.com Fri Feb 27 07:43:51 2009 From: mingsanma from gmail.com (Ming San Ma) Date: Fri Feb 27 12:07:22 2009 Subject: [Zbrafish] ISH + IHC Message-ID: <7577c5700902270443g26490079g35cdae5869132ab2@mail.gmail.com> Dear dr. Vayuputra, I came across your question regarding combined in situ hybridisation using anti-DIG-rhodamin as a detection method for your hybridisation, combined with fluorescent immunohistochemistry. I am trying the same now and encounter the same problems (no detection), while detection with anti-DIG AP and subsequent NBT-BCIP precipitation yields nice results. Have you succeeded in visualizing your probe fluorescently? If yes, I'd very much appreciate your advice. Thank you very much in advance, -- Ming-San Ma, MD Dept. of Neuroscience University Medical Center Groningen 9713 AV Groningen, The Netherlands +31-50-3632759 mingsanma@gmail.com www.mingsanma.com (under construction) -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090227/0a43816f/attachment.html From wclements from ucsd.edu Fri Feb 27 14:16:00 2009 From: wclements from ucsd.edu (Wilson Clements) Date: Fri Feb 27 14:19:29 2009 Subject: [Zbrafish] Fwd: Zbrafish Digest, Vol 45, Issue 16 References: <200902271703.n1RH3We12323@net.bio.net> Message-ID: <1FC64813-1C0C-416E-9AFF-0EC59FF4EF6A@ucsd.edu> Dear Jonathan, We contemplated the strategy you suggest, i.e. making a bicistronic transgene with cmlc2:gfp as a transgenesis reporter. We abandoned this strategy because we felt there was a high danger of crosstalk between the two promoter drivers. In your case, the danger would be cmlc2 driving your oncogene and mitfa driving eGfp. So if you don't care if your oncogene might get expressed in the heart, then I guess you wouldn't have to worry. I wasn't sure from your posting why you don't think you will be able to tell if the F1 are transgenic without cancer. I mean the mitfa promoter is active from late somitogenesis forward, so it seems to me you should be able to see fluorescence if you have a transgenic. Perhaps you are trying to do this by "transient transgenesis", which is to say looking at effects in the injected generation. If so, my main concern would be neoplastic effects downstream of ectopic expression. We see a lot of ectopic (especially somitic) expression of transgenes in the injected generation when using the Tol2 system, even when we get nice, clean expression of the transgene in the F1 stables. 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 Begin forwarded message: > From: zbrafish-request@oat.bio.indiana.edu > Date: February 27, 2009 9:03:32 AM PST > To: zbrafish@magpie.bio.indiana.edu > Subject: Zbrafish Digest, Vol 45, Issue 16 > Reply-To: zbrafish@oat.bio.indiana.edu > > 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. Zfish injection/integration control (Jonathan Schumacher) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Thu, 26 Feb 2009 11:10:10 -0800 (PST) > From: Jonathan Schumacher > Subject: [Zbrafish] Zfish injection/integration control > To: bionet-organisms-zebrafish@moderators.isc.org > Message-ID: > > Content-Type: text/plain; charset=ISO-8859-1 > > Greetings, > > I have designed a construct of MITFA driving my oncogene and > cytoplasmic-expressed EGFP, subsequently cloned into the pDest > Tol2pA. We suspect that it will take at least 8 weeks to see whether > the fish develop cancer and after setting up injections, I realized > that I have no injection/integration control. My question is would > you suggest recloning my construct into the pDest Tol2pA with cmlc2- > driven EGFP? I believe that this would lead to double-EGFP labeling > and am unclear if this would be problematic. Is there an alternative > strategy, like coinjecting my destination vector with a cherry-labeled > cmlc2 (or other)? > > Your thoughts & suggestions are greatly appreciated. > > Best, > > Jonathan > > > > ------------------------------ > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > > End of Zbrafish Digest, Vol 45, Issue 16 > **************************************** > -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090227/7bbbd9ab/attachment.html