From Roland.Dosch from unige.ch Tue Jan 13 05:48:30 2009 From: Roland.Dosch from unige.ch (DOSCH Roland) Date: Tue Jan 13 12:24:22 2009 Subject: [Zbrafish] SWISSZEBRA Meeting March 27, 2009 in Geneva, Switzerland Message-ID: Dear Colleagues, I wanted to remind you that the deadline for abstract submission for the 2nd Swisszebra meeting in Geneva, Switzerland on March 27, 2009, is coming up in three weeks (January 30th) and the max. number of participants is limited to 100. Registration is free, but required to login for abstract submission. For registration please check the meeting website, where you also find additional information about the meeting. http://www.swisszebra.ch In addition to the invited speakers (Hernan Lopez-Schier, Dirk Meyer, Steve Wilson) 6 short talks will be selected from the submitted poster abstracts. We look forward to seeing you in Geneva! Best regards, Bozena, Daniel, Stephan and Roland ______________________________________ Roland DOSCH Universit? de Gen?ve D?partement de Zoologie et Biologie Animale Sciences III 30, quai Ernest-Ansermet CH-1211 Gen?ve 4 SWITZERLAND Tel.: +41-(0)22-379-32 80 Fax.: +41-(0)22-379-33 40 http://www.biani.unige.ch/dosch !!!!! NEW EMAIL !!!!! roland.dosch@unige.ch -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090113/56389b1a/attachment.html From mena22787 from excite.com Tue Jan 13 17:35:21 2009 From: mena22787 from excite.com (Serena) Date: Tue Jan 13 17:49:49 2009 Subject: [Zbrafish] Re: cryopreservation service References: Message-ID: On Dec 29 2008, 3:37 pm, CryoDanio wrote: > Hello, > > I am interested in starting a business that would provide zebrafish > cryopreservation services, including sperm freezing, storage and > thawing for in vitro fertilization. I'm trying to get a feel for what > level of demand exists in the zebrafish community. > > As a former technician myself, I know the amount of time and resources > needed to perfect the skill of cryopreservation. It seems likely to > me that some labs would rather pay to have this done professionally > than incur the costs of learning it themselves. I would like to hear > other peoples' thoughts on the idea. > > Thanks in advance for your feedback! > > Joy We've been working on cryosections in my lab. Just out of curiosity, how much do you think you would charge per slide? Thanks. From cmoens from fhcrc.org Tue Jan 13 20:36:39 2009 From: cmoens from fhcrc.org (Cecilia Moens) Date: Wed Jan 14 13:02:07 2009 Subject: [Zbrafish] Submit TILLING requests Message-ID: <01D23B0B-7FF0-499B-AF00-78E965542E06@fhcrc.org> To the Members of the Zebrafish Community: The Zebrafish TILLING Consortium is pleased to announce the launch of a submission site for TILLING requests. TILLING is a method for finding unique ENU-induced mutations in specific genes in a library of mutagenized fish. This project is a joint effort of the Moens lab at the Fred Hutchinson Cancer Research Center, the Solnica-Krezel lab at Vanderbilt University, and the Postlethwait lab at the University of Oregon. The goal of the project is to generate and distribute loss-of- function mutations in genes of interest to the zebrafish community. Our labs have established large cryopreserved libraries of ENU- mutagenized zebrafish and have identified loss-of-function mutations in over 70 zebrafish genes to date. Our consortium-based approach will be to screen genes of interest sequentially at the three locations. This work is supported by a multi-PI NIH Grant HG002995-04, which provides salary support for TILLING personnel. The cost of TILLING consumables is passed on to the requesters. To learn more about the project and to submit your requests online, go to: http://www.fhcrc.org/tilling Yours truly, Cecilia Moens, Lila Solnica-Krezel and John Postlethwait. -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090113/89b789b7/attachment.html From marta.manzoni from gmail.com Wed Jan 14 12:43:32 2009 From: marta.manzoni from gmail.com (martam) Date: Wed Jan 14 13:02:34 2009 Subject: [Zbrafish] About morpholino heating before injection Message-ID: <5e0e58e6-35de-47dd-85a4-f028c7230142@f33g2000vbf.googlegroups.com> Every people I know use a different procedure to dissolve morpholinos before inject them. So... Which is the correct procedure to dilute morpholino and prepare the injection mixture? I dissolve morpholino in Danieu buffer 1mM and freeze it. First question: which is the best choice to store stock solution of MO, -20?C or -80?C?? Then, do I have to thaw the oligo before heating it to 65C for 10 minutes? Once cooled to room, dissolved and diluted to the final needed concentration, is it better to heat the injection mix again? Thanks a lot. From rburdine from Princeton.EDU Wed Jan 14 13:15:28 2009 From: rburdine from Princeton.EDU (Burdine, Rebecca D) Date: Wed Jan 14 13:19:00 2009 Subject: [Zbrafish] About morpholino heating before injection In-Reply-To: <5e0e58e6-35de-47dd-85a4-f028c7230142@f33g2000vbf.googlegroups.com> References: <5e0e58e6-35de-47dd-85a4-f028c7230142@f33g2000vbf.googlegroups.com> Message-ID: <04AF14C4EF3CF34682460FB1FDBA98C04A5D86@MBCLUSTER.pu.win.princeton.edu> Hi, We have used both Danieu's and water and haven't seen a difference in efficacy. We currently use water. In general, we make a very concentrated stock (50ug/ul) in water, aliquot into 5ul aliquots and store at -80C. When we thaw one to make dilutions for injections, we keep the stock and dilutions at 4 degrees from that point on. This is a relatively new procedure. We used to store everything at -80 and thaw when needed, but we found our MOs stopped working. Apparently others have seen this phenomena too, so Genetools recommended not freeze-thawing. Hopefully Paul will chime in here. I think it has something to do with the freeze thawing causing the MO to come out of solution? We make dilutions for injection into the standard KCl/Phenol red injection buffer. We do heat our dilutions to to 65 degree before injection. I don't think we typically heat the stock before making dilutions, but it can't hurt. I'd love to hear what other labs are doing. 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 > -----Original Message----- > From: zbrafish-bounces@oat.bio.indiana.edu [mailto:zbrafish- > bounces@oat.bio.indiana.edu] On Behalf Of martam > Sent: Wednesday, January 14, 2009 12:44 PM > To: bionet-organisms-zebrafish@moderators.isc.org > Subject: [Zbrafish] About morpholino heating before injection > > Every people I know use a different procedure to dissolve morpholinos > before inject them. So... > Which is the correct procedure to dilute morpholino and prepare the > injection mixture? > I dissolve morpholino in Danieu buffer 1mM and freeze it. > First question: which is the best choice to store stock solution of > MO, -20?C or -80?C?? > Then, do I have to thaw the oligo before heating it to 65C for 10 > minutes? > Once cooled to room, dissolved and diluted to the final needed > concentration, is it better to heat the injection mix again? > Thanks a lot. > > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish From jmoulton from gene-tools.com Wed Jan 14 15:19:31 2009 From: jmoulton from gene-tools.com (Jon D.Moulton) Date: Wed Jan 14 16:01:00 2009 Subject: [Fwd: RE: [Zbrafish] About morpholino heating before injection] Message-ID: <496E48D3.9010704@gene-tools.com> Hi all, This is Jon from Gene Tools. Here's our argument for water solution: (1) you can freeze-dry the oligo from a water solution, (2) you can prepare a sample for MALDI-TOF mass spectrometry from a water solution without ending up with a bunch of confounding salt-associated peaks in the spectrum, and (3) long ago at Antivirals Inc. (now AVI BioPharma Inc.) where Morpholinos were developed, Jim Summerton found that Morpholinos are more soluble in water than in buffers. Some Morpholino sequences are not soluble at concentrations much above 1 mM, which is why we recommend keeping Morpholinos in1mM stock solutions. If you made the solution in water and for some reason needed a higher concentration, you could lyophilize the oligo and redissolve it at higher concentration. Keeping a stock at a higher concentration encourages slow deposition of the oligo on the container walls, a process discussed below. When Morpholino activity drops over time, we know where it goes. The oligos associate with the walls of the container. There is a destructive test for this: pipet the fluid form the container and pipet in 0.l N HCl, shake, wait and test by UV spectrometry. Your missing 265 nm absorbance will reappear because the acid protonates the oligos on the wall and they dissolve back into solution. Morpholinos are very chemically and biochemically stable, so room temperature storage is an option to help keep the oligos in solution -- I suggest keeping them in a dark box with the vial closure wrapped in Parafilm to discourage microbial contamination. Never ice Morpholinos on the bench. If you have some that have decreased solution concentration, you can try autoclaving them (a few times -- I hesitate to recommend routine autoclaving, as we haven't tested for stability through many rounds of heat sterilization). Here is a link to the procedure for determining concentration by UV spectrometry. http://www.gene-tools.com/files/determining_concentration.pdf Let me know how I can help. Regards, - Jon Jon D. Moulton, Ph.D. GENE TOOLS, LLC jmoulton@gene-tools.com www.gene-tools.com (541) 929-7840 x1201 http://network.nature.com/group/morpholinos ----------------------------------------------------------------- Hi, We have used both Danieu's and water and haven't seen a difference in efficacy. We currently use water. In general, we make a very concentrated stock (50ug/ul) in water, aliquot into 5ul aliquots and store at -80C. When we thaw one to make dilutions for injections, we keep the stock and dilutions at 4 degrees from that point on. This is a relatively new procedure. We used to store everything at -80 and thaw when needed, but we found our MOs stopped working. Apparently others have seen this phenomena too, so Genetools recommended not freeze-thawing. Hopefully Paul will chime in here. I think it has something to do with the freeze thawing causing the MO to come out of solution? We make dilutions for injection into the standard KCl/Phenol red injection buffer. We do heat our dilutions to to 65 degree before injection. I don't think we typically heat the stock before making dilutions, but it can't hurt. I'd love to hear what other labs are doing. 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 > -----Original Message----- > From: zbrafish-bounces@oat.bio.indiana.edu [mailto:zbrafish- > bounces@oat.bio.indiana.edu] On Behalf Of martam > Sent: Wednesday, January 14, 2009 12:44 PM > To: bionet-organisms-zebrafish@moderators.isc.org > Subject: [Zbrafish] About morpholino heating before injection > > Every people I know use a different procedure to dissolve morpholinos > before inject them. So... > Which is the correct procedure to dilute morpholino and prepare the > injection mixture? > I dissolve morpholino in Danieu buffer 1mM and freeze it. > First question: which is the best choice to store stock solution of > MO, -20?C or -80?C?? > Then, do I have to thaw the oligo before heating it to 65C for 10 > minutes? > Once cooled to room, dissolved and diluted to the final needed > concentration, is it better to heat the injection mix again? > Thanks a lot. > > > _______________________________________________ > 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 smitra from abdn.ac.uk Thu Jan 15 08:32:02 2009 From: smitra from abdn.ac.uk (Mitra, Suman) Date: Thu Jan 15 12:37:27 2009 Subject: [Zbrafish] Zebrafish research in UK Message-ID: Hello all, I started my PhD at university of Aberdeen. I will need to FISH in zebrafish embryos. I wonder if any research lab in Europe will be able to help to do this experiment. Many thanks, suman The University of Aberdeen is a charity registered in Scotland, No SC013683. From jbennett from bio.umass.edu Fri Jan 16 11:03:01 2009 From: jbennett from bio.umass.edu (Judy Bennett) Date: Fri Jan 16 12:55:44 2009 Subject: [Zbrafish] cold weather shipping Message-ID: <336813CC-2CE7-402D-BAF6-A03FBFFF3E69@bio.umass.edu> Does anyone have a reliable protocol for (overnight) shipping adult fish and/or embryos in these frigid temps? Thanks Judy Bennett UMass, Amherst From Christian.Lawrence from childrens.harvard.edu Fri Jan 16 15:57:17 2009 From: Christian.Lawrence from childrens.harvard.edu (Lawrence, Christian) Date: Fri Jan 16 16:04:06 2009 Subject: [Zbrafish] cold weather shipping Message-ID: <54F197250C79354BA5A2D4584D8D9FDA3FAE51774F@CHEXCCRV1.CHBOSTON.ORG> D Lains at ZIRC does, I'm sure. 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 To: zbrafish@net.bio.net Sent: Fri Jan 16 11:03:01 2009 Subject: [Zbrafish] cold weather shipping Does anyone have a reliable protocol for (overnight) shipping adult fish and/or embryos in these frigid temps? Thanks Judy Bennett UMass, Amherst _______________________________________________ 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 bmebane from mbl.edu Fri Jan 16 16:35:00 2009 From: bmebane from mbl.edu (Bill Mebane) Date: Fri Jan 16 16:54:02 2009 Subject: [Zbrafish] cold weather shipping In-Reply-To: <54F197250C79354BA5A2D4584D8D9FDA3FAE51774F@CHEXCCRV1.CHBOSTON.ORG> References: <54F197250C79354BA5A2D4584D8D9FDA3FAE51774F@CHEXCCRV1.CHBOSTON.ORG> Message-ID: Judy - we are having some luck shipping embryos overnight using "Hot Hands" (disposable chemically activated hand warmers sold at fishing and hunting stores for appx $1/ea) There are a variety of manuf. but "Hot Hands" seems to be the best. The box should be one of the molded 2" thick styrofoam boxes...not the 3/4" foam boxes. The hand warmer has an adhesive side that we stick to the inside of the shipping box so the hand warmer is not in direct contact with the water bag. When the weather is really cold (<10F) 2-3 warmers are used - hint...it is possible to get a bad batch of these warmers so test one or two before you buy a box off the self. Bill At 3:57 PM -0500 1/16/09, Lawrence, Christian wrote: >D Lains at ZIRC does, I'm sure. > >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 > >To: zbrafish@net.bio.net >Sent: Fri Jan 16 11:03:01 2009 >Subject: [Zbrafish] cold weather shipping > >Does anyone have a reliable protocol for (overnight) shipping adult >fish and/or embryos in these frigid temps? > >Thanks >Judy Bennett >UMass, Amherst > >_______________________________________________ >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. > > > >_______________________________________________ >Zbrafish mailing list >Zbrafish@net.bio.net >http://www.bio.net/biomail/listinfo/zbrafish -- Superintendent, Aquaculture Engineering Division Marine Resources Center, Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543-1015 phone: (508) 289-7683 fax: (508) 289-7900 email: bmebane@mbl.edu http://www.mbl.edu (MBL main website) http://www.mbl.edu/haiti (Sustainable Aquaculture Project website) From jeedward from yahoo.com Fri Jan 16 17:04:12 2009 From: jeedward from yahoo.com (John Edward) Date: Fri Jan 16 17:15:31 2009 Subject: [Zbrafish] BCBGC-09 final call for papers Message-ID: <338974.84850.qm@web45915.mail.sp1.yahoo.com> BCBGC-09 final call for papers ? The 2009 International Conference on Bioinformatics, Computational Biology, Genomics and Chemoinformatics (BCBGC-09) (website: http://www.PromoteResearch.org ) 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/20090116/8a3760ef/attachment.html From mary_georger from URMC.Rochester.edu Fri Jan 16 17:06:37 2009 From: mary_georger from URMC.Rochester.edu (Mary Georger) Date: Fri Jan 16 17:15:54 2009 Subject: [Zbrafish] cold weather shipping In-Reply-To: Message-ID: Walgreen's also sells them by the box up here in the Northeast :) On 1/16/09 4:35 PM, "Bill Mebane" wrote: > Judy - we are having some luck shipping embryos overnight using "Hot > Hands" (disposable chemically activated hand warmers sold at fishing > and hunting stores for appx $1/ea) There are a variety of manuf. but > "Hot Hands" seems to be the best. The box should be one of the molded > 2" thick styrofoam boxes...not the 3/4" foam boxes. The hand warmer > has an adhesive side that we stick to the inside of the shipping box > so the hand warmer is not in direct contact with the water bag. When > the weather is really cold (<10F) 2-3 warmers are used - hint...it is > possible to get a bad batch of these warmers so test one or two > before you buy a box off the self. > Bill > > At 3:57 PM -0500 1/16/09, Lawrence, Christian wrote: >> D Lains at ZIRC does, I'm sure. >> >> 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 >> >> To: zbrafish@net.bio.net >> Sent: Fri Jan 16 11:03:01 2009 >> Subject: [Zbrafish] cold weather shipping >> >> Does anyone have a reliable protocol for (overnight) shipping adult >> fish and/or embryos in these frigid temps? >> >> Thanks >> Judy Bennett >> UMass, Amherst >> >> _______________________________________________ >> 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. >> >> >> >> _______________________________________________ >> Zbrafish mailing list >> Zbrafish@net.bio.net >> http://www.bio.net/biomail/listinfo/zbrafish > From bdschyth from gmail.com Sat Jan 17 06:32:00 2009 From: bdschyth from gmail.com (bdschyth) Date: Tue Jan 20 12:53:30 2009 Subject: [Zbrafish] cell line transfection Message-ID: <51977833-8d4a-4f6d-b8b9-2ad0476f4a69@v39g2000pro.googlegroups.com> To my experience and according to litterature RNA/DNA transfection of fish cell cultures is often difficult (low transfection %) so I?m seeking good protocols for fish cell cultures / easy transfectable cell cultures all the best From jbennett from bio.umass.edu Mon Jan 19 09:44:17 2009 From: jbennett from bio.umass.edu (Judy Bennett) Date: Tue Jan 20 12:54:44 2009 Subject: [Zbrafish] cold weather shipping In-Reply-To: References: <54F197250C79354BA5A2D4584D8D9FDA3FAE51774F@CHEXCCRV1.CHBOSTON.ORG> Message-ID: <9B6E8D91-B4B9-4EFD-B029-369AED0F47FC@bio.umass.edu> I think we were going to possibly go this route, but were wondering if they would last long enough... Thanks for the response. Judy On Jan 16, 2009, at 4:35 PM, Bill Mebane wrote: > Judy - we are having some luck shipping embryos overnight using > "Hot Hands" (disposable chemically activated hand warmers sold at > fishing and hunting stores for appx $1/ea) There are a variety of > manuf. but "Hot Hands" seems to be the best. The box should be one > of the molded 2" thick styrofoam boxes...not the 3/4" foam boxes. > The hand warmer has an adhesive side that we stick to the inside of > the shipping box so the hand warmer is not in direct contact with > the water bag. When the weather is really cold (<10F) 2-3 warmers > are used - hint...it is possible to get a bad batch of these > warmers so test one or two before you buy a box off the self. > Bill > > At 3:57 PM -0500 1/16/09, Lawrence, Christian wrote: >> D Lains at ZIRC does, I'm sure. >> >> 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 > bounces@oat.bio.indiana.edu> >> To: zbrafish@net.bio.net >> Sent: Fri Jan 16 11:03:01 2009 >> Subject: [Zbrafish] cold weather shipping >> >> Does anyone have a reliable protocol for (overnight) shipping >> adult fish and/or embryos in these frigid temps? >> >> Thanks >> Judy Bennett >> UMass, Amherst >> >> _______________________________________________ >> 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. >> >> >> >> _______________________________________________ >> Zbrafish mailing list >> Zbrafish@net.bio.net >> http://www.bio.net/biomail/listinfo/zbrafish > > > -- > Superintendent, Aquaculture Engineering Division > Marine Resources Center, Marine Biological Laboratory > 7 MBL Street > Woods Hole, MA 02543-1015 > phone: (508) 289-7683 > fax: (508) 289-7900 > email: bmebane@mbl.edu > http://www.mbl.edu (MBL main website) > http://www.mbl.edu/haiti (Sustainable Aquaculture Project website) From waldrons from u.washington.edu Fri Jan 16 13:06:14 2009 From: waldrons from u.washington.edu (Steven Waldron) Date: Tue Jan 20 12:55:24 2009 Subject: [Zbrafish] cold weather shipping In-Reply-To: <336813CC-2CE7-402D-BAF6-A03FBFFF3E69@bio.umass.edu> References: <336813CC-2CE7-402D-BAF6-A03FBFFF3E69@bio.umass.edu> Message-ID: <4970CC96.7050207@u.washington.edu> Hi Judy, You'll need to pack the fish in a small thick-walled styrofoam shipping container nestled within a cardboard box filled with packing peanuts. It is essential to pack the styrofoam peanut layer with a few heat packs- the kind used as hand-warmers for cold-weather sports will do. If the fish are going really long-distance you may have to invest in heat packs that are specifically designed for shipping livestock and maintain heat for a longer duration. You can find these at aquabid.com. best, Steve p.s. make sure the heat packs do not come into direct contact with the container holding the fish...ironically, direct contact with the heat packs could fry your fish even if the outside temperature is frigid. Judy Bennett wrote: > Does anyone have a reliable protocol for (overnight) shipping adult > fish and/or embryos in these frigid temps? > > Thanks > Judy Bennett > UMass, Amherst > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish From weibinz from med.umich.edu Tue Jan 20 12:49:08 2009 From: weibinz from med.umich.edu (Weibin Zhou) Date: Tue Jan 20 12:56:04 2009 Subject: [Zbrafish] in situ and engrailed staining Message-ID: <4975C843.8167.0008.0@med.umich.edu> Hi, I am wondering whether someone would be willing to share his/her experience doing in situ and immunolabeling with monoclonal antibody 4D9 (engrailed). I have tried to do this double labeling but it appears that the antibody did not work well after the in situ procedure. I appreciate very much if someone would supply any suggestions or comments. Thank you very much. weibin ********************************************************** Electronic Mail is not secure, may not be read every day, and should not be used for urgent or sensitive issues From Christian.Lawrence from childrens.harvard.edu Tue Jan 20 14:23:29 2009 From: Christian.Lawrence from childrens.harvard.edu (Lawrence, Christian) Date: Tue Jan 20 14:28:22 2009 Subject: [Zbrafish] cold weather shipping In-Reply-To: <4970CC96.7050207@u.washington.edu> Message-ID: It's probably worth mentioning that as long as the eggs are properly packed (bleached, 1 dpf at ~ 1 embryo/ml in sealed conical tubes or tissue culture flasks wrapped in bubble wrap) inside sealed thick-walled styrofoam containers, no heating packs are necessary IF the fish are received within 24-36 hours after packing. Beyond this, it can get a bit dicey. But we routinely (on a nearly weekly basis) ship domestically without heat packs during cold weather months with no reported problems. The embryos are remarkably tolerant of temperature swings. This isn't too surprising given that zebrafish are among the most eurythermal fish species on record. A useful little study would be to track temperatures inside sealed packing containers shipped to various locations at different times of year. Perhaps ZIRC has this kind of data? 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 On 1/16/09 1:06 PM, "Steven Waldron" wrote: Hi Judy, You'll need to pack the fish in a small thick-walled styrofoam shipping container nestled within a cardboard box filled with packing peanuts. It is essential to pack the styrofoam peanut layer with a few heat packs- the kind used as hand-warmers for cold-weather sports will do. If the fish are going really long-distance you may have to invest in heat packs that are specifically designed for shipping livestock and maintain heat for a longer duration. You can find these at aquabid.com. best, Steve p.s. make sure the heat packs do not come into direct contact with the container holding the fish...ironically, direct contact with the heat packs could fry your fish even if the outside temperature is frigid. Judy Bennett wrote: > Does anyone have a reliable protocol for (overnight) shipping adult > fish and/or embryos in these frigid temps? > > Thanks > Judy Bennett > UMass, Amherst > > _______________________________________________ > 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 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 david from zebrafish.org Wed Jan 21 13:33:34 2009 From: david from zebrafish.org (David Lains) Date: Wed Jan 21 13:36:25 2009 Subject: [Zbrafish] cold weather shipping In-Reply-To: <9B6E8D91-B4B9-4EFD-B029-369AED0F47FC@bio.umass.edu> References: <54F197250C79354BA5A2D4584D8D9FDA3FAE51774F@CHEXCCRV1.CHBOSTON.ORG> <9B6E8D91-B4B9-4EFD-B029-369AED0F47FC@bio.umass.edu> Message-ID: <05a401c97bf6$c7baa900$572ffb00$@org> Hello Judy I use 35 hour packs for domestic and 60 hour packs for international shipments. They usually last about 48 and 72 hours respectively but they are less reliable at the ends of their service. http://www.theenergykit.com/Products7.asp The packs are highly reliable but I always start them with oxygen in a fish bag to make sure they start producing heat. To protect the heat packs from touching the flasks or bags I put them in a bubble wrap envelope with the corners cut off and a slit down the center to keep them from suffocating. For embryos I put the pad in the bottom of the box with about an inch or two of peanuts between the flask and pad. Make sure the peanuts are packed tight or the flask can settle down on to the pad due to the vibrations in transit. For the adults I put them in the bubble wrap and tape them to the inside of the lid. If you are shipping adults and embryos together make sure the flasks touch the bag below the water level. This keeps the embryos warmer by making them part of a larger thermal mass. If I'm sending embryos only to a very cold location like Fairbanks last week I'll often put a liter or two of water into a fish bag just to help moderate the temperature. 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 -----Original Message----- From: zbrafish-bounces@oat.bio.indiana.edu [mailto:zbrafish-bounces@oat.bio.indiana.edu] On Behalf Of Judy Bennett Sent: Monday, January 19, 2009 6:44 AM To: Bill Mebane Cc: 'zbrafish@magpie.bio.indiana.edu'; Lawrence, Christian Subject: Re: [Zbrafish] cold weather shipping I think we were going to possibly go this route, but were wondering if they would last long enough... Thanks for the response. Judy On Jan 16, 2009, at 4:35 PM, Bill Mebane wrote: > Judy - we are having some luck shipping embryos overnight using > "Hot Hands" (disposable chemically activated hand warmers sold at > fishing and hunting stores for appx $1/ea) There are a variety of > manuf. but "Hot Hands" seems to be the best. The box should be one > of the molded 2" thick styrofoam boxes...not the 3/4" foam boxes. > The hand warmer has an adhesive side that we stick to the inside of > the shipping box so the hand warmer is not in direct contact with > the water bag. When the weather is really cold (<10F) 2-3 warmers > are used - hint...it is possible to get a bad batch of these > warmers so test one or two before you buy a box off the self. > Bill > > At 3:57 PM -0500 1/16/09, Lawrence, Christian wrote: >> D Lains at ZIRC does, I'm sure. >> >> 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 > bounces@oat.bio.indiana.edu> >> To: zbrafish@net.bio.net >> Sent: Fri Jan 16 11:03:01 2009 >> Subject: [Zbrafish] cold weather shipping >> >> Does anyone have a reliable protocol for (overnight) shipping >> adult fish and/or embryos in these frigid temps? >> >> Thanks >> Judy Bennett >> UMass, Amherst >> >> _______________________________________________ >> 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. >> >> >> >> _______________________________________________ >> Zbrafish mailing list >> Zbrafish@net.bio.net >> http://www.bio.net/biomail/listinfo/zbrafish > > > -- > Superintendent, Aquaculture Engineering Division > Marine Resources Center, Marine Biological Laboratory > 7 MBL Street > Woods Hole, MA 02543-1015 > phone: (508) 289-7683 > fax: (508) 289-7900 > email: bmebane@mbl.edu > http://www.mbl.edu (MBL main website) > http://www.mbl.edu/haiti (Sustainable Aquaculture Project website) _______________________________________________ Zbrafish mailing list Zbrafish@net.bio.net http://www.bio.net/biomail/listinfo/zbrafish From chi-bin.chien from neuro.utah.edu Wed Jan 21 17:56:34 2009 From: chi-bin.chien from neuro.utah.edu (Chi-Bin Chien) Date: Wed Jan 21 18:16:59 2009 Subject: [Zbrafish] GFP and in situ In-Reply-To: <20080801134657.25997h14wd85hoo4@web.mail.umich.edu> References: <20080801134657.25997h14wd85hoo4@web.mail.umich.edu> Message-ID: <16366DB6-C050-43DB-B38C-170D80C34D9F@neuro.utah.edu> Weibin-- No, I don't think this is possible. GFP fluorescence is delicate, in situs are harsh. However, GFP antigenicity can survive quite well, and we routinely antibody stain for GFP after in situs. Chi-Bin Chien On Aug 1, 2008, at 11:46 AM, Weibin Zhou wrote: > Hi, > I am wondering whether it is possible to preserve GFP fluorescence > after in situ hybridization (whole-mount or sectioned). If you > happen to have a protocol that works, please let me know. Thank you > very much. > > Weibin > > > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > From jaschumac from gmail.com Tue Jan 27 09:56:31 2009 From: jaschumac from gmail.com (Jonathan Schumacher) Date: Tue Jan 27 12:51:55 2009 Subject: [Zbrafish] Construct/transposase [ ] Message-ID: <99288b0b-74ac-4969-89f5-5b9fa145b031@g1g2000pra.googlegroups.com> 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: schumaj@aruplab.com From chi-bin.chien from neuro.utah.edu Tue Jan 27 13:06:34 2009 From: chi-bin.chien from neuro.utah.edu (Chi-Bin Chien) Date: Tue Jan 27 13:22:41 2009 Subject: [Zbrafish] Construct/transposase [ ] In-Reply-To: <99288b0b-74ac-4969-89f5-5b9fa145b031@g1g2000pra.googlegroups.com> References: <99288b0b-74ac-4969-89f5-5b9fa145b031@g1g2000pra.googlegroups.com> Message-ID: <796E1B05-4EAC-4ADA-8A67-CAE2679FA609@neuro.utah.edu> Hi Jonathan, Those concentrations are reasonable, assuming that you are injecting about 1 nl/embryo. If you read any of the original papers on the Tol2 transposon you will find details. For instance, look at our Tol2kit paper: KM Kwan et al. (2007) The Tol2kit: a multisite Gateway-based construction kit for Tol2 transposon transgenesis constructs. Developmental Dynamics, 236:3088-99. Or, read the Tol2kit wiki: http://chien.neuro.utah.edu/Tol2kitwiki/ --Chi-Bin Chien On Jan 27, 2009, at 7:56 AM, 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: schumaj@aruplab.com > > _______________________________________________ > 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/20090127/488b0308/attachment.html From jmurtha from umich.edu Thu Jan 29 11:20:42 2009 From: jmurtha from umich.edu (jmurtha@umich.edu) Date: Thu Jan 29 11:22:38 2009 Subject: [Zbrafish] gill biopsy Message-ID: hello, has anyone done gill biopsies on zebrafish? is it possible to do this as a non-terminal procedure? any info would be appreciated. thanks, jill From jaschumac from gmail.com Thu Jan 29 13:42:54 2009 From: jaschumac from gmail.com (Jonathan Schumacher) Date: Thu Jan 29 13:46:21 2009 Subject: [Zbrafish] making gels for embryo injections Message-ID: Greetings, In "The Zebrafish Book", I see a really nice description of a device to hold zebrafish embryos during microinjection (pg. 5.2). I'd like to make gels using this plastic "slot maker", but don't know where to find such a device. I've been using standard vertical wells, but it would be nice to have 1 vertical wall to rest the embryo against & 1 sloped wall to inject from. Any idea where to purchase this "slot maker"? Kindest regards, Jonathan From anandc from missouri.edu Fri Jan 30 09:45:35 2009 From: anandc from missouri.edu (Anand) Date: Fri Jan 30 12:30:09 2009 Subject: [Zbrafish] Re: making gels for embryo injections References: Message-ID: Hi Jonathan, We had ours made in the workshop at MPI Tubingen. Most university workshops (esp. in Physics) should be able to make them from plexiglass blocks. Best wishes, Anand Chandrasekhar On Jan 29, 12:42?pm, Jonathan Schumacher wrote: > Greetings, > > In "The Zebrafish Book", I see a really nice description of a device > to hold zebrafish embryos during microinjection (pg. 5.2). ?I'd like > to make gels using this plastic "slot maker", but don't know where to > find such a device. ?I've been using standard vertical wells, but it > would be nice to have 1 vertical wall to rest the embryo against & 1 > sloped wall to inject from. ?Any idea where to purchase this "slot > maker"? > > Kindest regards, > > Jonathan From emond.4 from osu.edu Fri Jan 30 12:18:12 2009 From: emond.4 from osu.edu (Michelle Emond) Date: Fri Jan 30 12:30:31 2009 Subject: [Zbrafish] Re: Injection Molds Message-ID: <49833654.9040608@osu.edu> Hi, We first acquired the molds at a meeting a long time ago, but they became impossible to find for a long time. However, a company has started providing them again. They sell a variety of mold styles for $25 each: Adaptive Science Tools http://adaptivesciencetools.com/index.html (774) 239-6133 From fritsa01 from gettysburg.edu Fri Jan 30 00:22:37 2009 From: fritsa01 from gettysburg.edu (FritzXC23) Date: Fri Jan 30 12:30:58 2009 Subject: [Zbrafish] RALDH Protein Assay Message-ID: <16653747-aa5f-4e40-af0b-0d858b78740c@f18g2000vbf.googlegroups.com> Hi! My name is Sarah and I am undergraduate Biochemistry student at Gettysburg College. I am currently conducting an independent research experiment investigating the effects of vitamin A deficiency on eye development in zebrafish. To make the embryos vitamin A deficient I am dosing them with various concentrations of disulphiram and DEAB (two retinaldehyde dehydrogenase inhibitors). I was wondering if anyone could give me advice on a protein assay that I could potentially perform that would allow me to measure RALDH activity levels in the dosed fish. I found an article that described a BioRad assay to measure protein concentration followed by a simple spectrophotometric assay at 350nm. The tissue preparation described for these two assays in regards to RALDH activity, however, was from a human source. Has anyone performed or heard of these assays and know how to prepare zebrafish embryos for them? Or does anyone have a better suggestion for an assay that I could reasonably perform? Thanks for your help! Sarah From jc.achenbach from nrc.ca Fri Jan 30 09:12:55 2009 From: jc.achenbach from nrc.ca (J.C.) Date: Fri Jan 30 12:31:13 2009 Subject: [Zbrafish] Re: making gels for embryo injections References: Message-ID: Hi, we've purchased ours from Adaptive Science tools from Worchester, MA. Model TU-1 is the type you describe but they have a couple other options as well. http://www.adaptivesciencetools.com/ happy injecting, J.C. From lhqkey from gmail.com Fri Jan 30 09:39:06 2009 From: lhqkey from gmail.com (Haiqiong) Date: Fri Jan 30 12:31:32 2009 Subject: [Zbrafish] Re: making gels for embryo injections References: Message-ID: <97955430-1b4f-4923-90e8-cb8542383561@40g2000prx.googlegroups.com> On Jan 29, 2:42?pm, Jonathan Schumacher wrote: Hi Jonathan, I just use several glass microscope slides stick together with transparent nail polish (so that it is thick enough), and tilt it in the petri dish. It only makes one row (for holding embryos) per petri dish. It works very well for me. Maybe you can give it a try. Haiqiong Fekete Lab Purdue University > Greetings, > > In "The Zebrafish Book", I see a really nice description of a device > to hold zebrafish embryos during microinjection (pg. 5.2). ?I'd like > to make gels using this plastic "slot maker", but don't know where to > find such a device. ?I've been using standard vertical wells, but it > would be nice to have 1 vertical wall to rest the embryo against & 1 > sloped wall to inject from. ?Any idea where to purchase this "slot > maker"? > > Kindest regards, > > Jonathan From cmoens from fhcrc.org Fri Jan 30 13:23:56 2009 From: cmoens from fhcrc.org (Cecilia Moens) Date: Fri Jan 30 13:39:59 2009 Subject: [Zbrafish] making gels for embryo injections (Jonathan Schumacher) Message-ID: <1903870E-1D47-42A1-BC59-9A5A4705D271@fhcrc.org> 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 -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090130/952faf72/attachment.html