From michael.lardelli from adelaide.edu.au Fri May 1 00:31:22 2009 From: michael.lardelli from adelaide.edu.au (Michael Lardelli) Date: Fri May 1 11:25:56 2009 Subject: [Zbrafish] Analgesia in Fish Message-ID: <001b01c9ca1e$104d97c0$526b7f81@ad.adelaide.edu.au> I would like to direct the attention of zebrafish researchers to the following interesting conference paper: http://www.royalsociety.org.nz/includes/download.aspx?ID=101797 "Pain" and analgesia in fish: What we know, what we do not know, and what we need to know, before using analgesics in fish by Professor E. Don Stevens Department of Biomedical Sciences Atlantic Veterinary College University of Prince Edward Island Charlottetown, PE Canada C1A 4P3 Regards, Michael Michael Lardelli Zebrafish Genetics Laboratory School of Molecular and Biomedical Science The University of Adelaide, AUSTRALIA 5005 Ph : +61 8 8303 3212 Fax : +61 8 8303 4362 e-mail: michael.lardelli@adelaide.edu.au CRICOS Provider Number 00123M ----------------------------------------------------------- This email message is intended only for the addressee(s) and contains information that may be confidential and/or copyright. If you are not the intended recipient please notify the sender by reply email and immediately delete this email. Use, disclosure or reproduction of this email by anyone other than the intended recipient(s) is strictly prohibited. No representation is made that this email or any attachments are free of viruses. Virus scanning is recommended and is the responsibility of the recipient. From ilovequestions99 from gmail.com Mon May 4 11:19:12 2009 From: ilovequestions99 from gmail.com (Wei Xia) Date: Mon May 4 12:53:55 2009 Subject: [Zbrafish] Help with Transgenic constructi Message-ID: Hi everyone, I'm trying to study the regulatory sequences of a specific promoter. So I will fuse the promoter to a LacZ reporter to generate a transgenic zebrafish and see If It can drive expression of the LacZ in specifics areas of the zebrafish brain. I'm just wondering If in my construction I have to include the ATG (translation start site) of the gene's promoter, or If it will works only with the transcription start site, knowing that the LacZ Reporter has a start codon? Thank's for your help! -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090504/f54abff0/attachment.html From michael.lardelli from adelaide.edu.au Sun May 3 19:23:22 2009 From: michael.lardelli from adelaide.edu.au (Michael Lardelli) Date: Mon May 4 12:54:18 2009 Subject: [Zbrafish] Analgesia in Fish In-Reply-To: <5D5D2B2F-E5E5-4273-A809-6E68B883BF1A@neuro.utah.edu> References: <001b01c9ca1e$104d97c0$526b7f81@ad.adelaide.edu.au> <5D5D2B2F-E5E5-4273-A809-6E68B883BF1A@neuro.utah.edu> Message-ID: <005c01c9cc4e$8875b2a0$526b7f81@ad.adelaide.edu.au> Dear Zebrafish Researchers, I have been informed that the method for citation of Don Stevens' paper on analgesia in fish is: Don E. Stevens, "Pain" and analgesia in fish: What we know, what we don't know, and what we need to know before using analgesics in fish. Pp. 115-124 in: Proceedings of 2008 ANZCCART Conference, Auckland, New Zealand. Regards, Michael > > I would like to direct the attention of zebrafish researchers to the > following interesting conference paper: > > http://www.royalsociety.org.nz/includes/download.aspx?ID=101797 > > "Pain" and analgesia in fish: What we know, what we do not know, and > what we need to know, before using analgesics in fish > > by > > Professor E. Don Stevens > Department of Biomedical Sciences > Atlantic Veterinary College > University of Prince Edward Island > Charlottetown, PE Canada C1A 4P3 > > > Regards, > > Michael > > > Michael Lardelli > Zebrafish Genetics Laboratory > School of Molecular and Biomedical Science The University of Adelaide, > AUSTRALIA 5005 > Ph : +61 8 8303 3212 > Fax : +61 8 8303 4362 > e-mail: michael.lardelli@adelaide.edu.au CRICOS Provider Number 00123M > ----------------------------------------------------------- > This email message is intended only for the addressee(s) and contains > information that may be confidential and/or copyright. If you are not > the intended recipient please notify the sender by reply email and > immediately delete this email. Use, disclosure or reproduction of this > email by anyone other than the intended recipient(s) is strictly > prohibited. No representation is made that this email or any > attachments are free of viruses. Virus scanning is recommended and is > the responsibility of the recipient. > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish From zoltan from zebrafish.org Wed May 6 12:48:14 2009 From: zoltan from zebrafish.org (Zoltan) Date: Wed May 6 14:24:51 2009 Subject: [Zbrafish] ZIRC & UCL Nursery Workshop in Rome Message-ID: <621b51a3-1c5e-4d47-ab4d-62b7c18b66fa@u39g2000pru.googlegroups.com> Dear all, ZIRC and UCL are planning a Husbandry Workshop with emphasis on Larval Care and Nursery Protocols during the '6th European Zebrafish, Genetics and Development Meeting’, in Rome, Italy (15-19th July). If you have interesting ideas, protocols, problems, tips, tricks, or concerns that you think we should share and discuss at the workshop, please contact us (see below). Specifically, if you would like to give a 10-minute presentation on your own nursery practices, please send us an abstract. We are trying to cover the topics below. However, additional suggestions for interesting, helpful nursery topics are very welcome! Any single one of these, or several of these (and/or additional) issues can be addressed in your presentation (10 minute max!): - Pre-nursery embryo care - Nursery work protocols and organization - Larval feeding and (live) food - Nursery trouble-shooting - Nursery Equipment - Tips & Tricks Please send your ideas, suggestions, and proposals to Zoltan Varga zoltan@zebrafish.org or Carole Wilson carole.wilson@ucl.ac.uk We are very much looking forward to seeing you at the workshop! Yours, Carole and Zoltan From sophielouwette from hotmail.com Tue May 12 03:00:30 2009 From: sophielouwette from hotmail.com (Sophie Louwette) Date: Tue May 12 11:02:43 2009 Subject: [Zbrafish] ratio males/females Message-ID: Dear all, Lately we are having problems with raising our zebrafish. The survival rate is fine (70%-80%) but the majority of them are males. We keep our fish on 26?C and not too crowded. I was wondering if there are correlations between temperature and density and the ratio of females/males. Maybe you can give us some advice on this issue. Best regards, Sophie _________________________________________________________________ Je hele online leven op ??n stek met Windows Live http://www.microsoft.com/belux/nl/windows/windowslive/default.aspx -------------- next part -------------- An HTML attachment was scrubbed... URL: http://www.bio.net/bionet/mm/zbrafish/attachments/20090512/7b1cd166/attachment.html From dgw5079 from u.washington.edu Tue May 12 17:36:15 2009 From: dgw5079 from u.washington.edu (David G. White) Date: Tue May 12 17:39:10 2009 Subject: [Zbrafish] ratio males/females In-Reply-To: Message-ID: What are you D.O. levels? See link below. http://www.usatoday.com/tech/science/2006-03-29-dead-fish-zones_x.htm David G White Research Coordinator H225 Zebrafish Lab University of Washington Department of Biological Structure HSB G514 Box 357420 1959 NE Pacific Street Seattle, WA 98195-7420 Tel. 206-685-7512 FAX 206-543-1524 On Tue, 12 May 2009, Sophie Louwette wrote: > > Dear all, > > > > Lately we are having problems with raising our zebrafish. The survival rate is fine (70%-80%) but the majority of them are males. We keep our fish on 26?C and not too crowded. I was wondering if there are correlations between temperature and density and the ratio of females/males. Maybe you can give us some advice on this issue. > > > > Best regards, > > Sophie > > > > > > _________________________________________________________________ > Je hele online leven op ??n stek met Windows Live > http://www.microsoft.com/belux/nl/windows/windowslive/default.aspx From busquet.francois from gmail.com Wed May 13 02:23:25 2009 From: busquet.francois from gmail.com (=?ISO-8859-1?Q?fran=E7ois?=) Date: Wed May 13 11:20:10 2009 Subject: [Zbrafish] Re: ratio males/females References: Message-ID: On May 12, 10:00 am, Sophie Louwette wrote: > Dear all, > > Lately we are having problems with raising our zebrafish. The survival rate is fine (70%-80%) but the majority of them are males. We keep our fish on 26°C and not too crowded. I was wondering if there are correlations between temperature and density and the ratio of females/males. Maybe you can give us some advice on this issue. > > Best regards, > > Sophie > > _________________________________________________________________ > Je hele online leven op één stek met Windows Livehttp://www.microsoft.com/belux/nl/windows/windowslive/default.aspx you can have a look at this paper: 2005 Animal Behaviour , 69, 1317–1323 Male territoriality mediates density and sex ratio effects on oviposition in the zebrafish, Danio rerio Rowena Spence, and Carl Smith Francois From Christian.Lawrence from childrens.harvard.edu Wed May 13 12:14:35 2009 From: Christian.Lawrence from childrens.harvard.edu (Lawrence, Christian) Date: Wed May 13 12:15:26 2009 Subject: [Zbrafish] ratio males/females In-Reply-To: Message-ID: There are a number of factors that influence sexual differentiation in zebrafish. Temperature is most often implicated but is unlikely to be the environmental factor at work in fish facilities, since published data indicates that masculinizing temperatures are in the mid to low 30s. Food consumption, mediated by densities, is more likely to affect lab stocks, although if there is a bias caused by this, it is more likely to be a female one, since in practice it is much easier to overfeed and the scenario (female bias) is more in line with the potential underlying mechanism (estrogen content in feeds). The other well known environmental factor known to impact sexual differentiation is dissolved oxygen. Very low do2 (hypoxic conditions) can exert masculinizing effects. Certainly this is conceivable on nursery racks especially but given reports on this effect in the literature, one would also expect to see high mortality, with a strong male bias in survivors. This doesn't seem to be your pattern, either. It may not be environmental. Sex ratios can be heritable in zebrafish. Certain crosses will sometimes yield a higher proportion of one sex over the other. This reasons for this are not well understood but this seems to tend to crop up in facilities where stocks are maintained in small, closed populations for extended number of generations. The sex ratio effect seems to be most pronounced when fish from one closed population are crossed to another. Again, in the literature, and in our hands, the bias seems to typically be female, but it may work in the other direction. Quite conveniently, it may also be a combination of the above, or it could even be some synthetic masculining agent leaching into the water. Such are the joys of working with a teleost fish with a yet to be determined and likely polygenic sex determining system. No easy answers! 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 5/12/09 4:00 AM, "Sophie Louwette" wrote: Dear all, Lately we are having problems with raising our zebrafish. The survival rate is fine (70%-80%) but the majority of them are males. We keep our fish on 26?C and not too crowded. I was wondering if there are correlations between temperature and density and the ratio of females/males. Maybe you can give us some advice on this issue. Best regards, Sophie ________________________________ Je hele online leven op ??n stek? Ontdek Windows Live! From caroline.parkin from sheffield.ac.uk Wed May 20 06:31:03 2009 From: caroline.parkin from sheffield.ac.uk (Caroline Parkin) Date: Wed May 20 10:45:33 2009 Subject: [Zbrafish] How many scientists and or labs use ZF? Message-ID: Hello all, I'd like to know if anyone as an idea of how many labs there are in the world using zebrafish for research, or how many researchers there are? I know this is likely to be a bit vague because of collaborators etc..but an order of magnitude that we can all agree on would be great! And if anyone can estimate how many fish or fish lines even there are then that would be fantastic! Many thanks, Caroline From rburdine from Princeton.EDU Wed May 20 12:35:56 2009 From: rburdine from Princeton.EDU (Burdine, Rebecca D) Date: Wed May 20 12:37:26 2009 Subject: [Zbrafish] How many scientists and or labs use ZF? In-Reply-To: References: Message-ID: <790DAC09623D0A47ACDB126E3C52D8B61DC217@MBCLUSTER.pu.win.princeton.edu> I think your best bet would be to contact ZFIN and ask how many labs are registered with the network. It won't get absolutely everyone who works with fish, but it will get the major players. 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 Caroline Parkin > Sent: Wednesday, May 20, 2009 7:31 AM > To: zbrafish@magpie.bio.indiana.edu > Subject: [Zbrafish] How many scientists and or labs use ZF? > > Hello all, > > I'd like to know if anyone as an idea of how many labs there are in > the world using zebrafish for research, or how many researchers there > are? > > I know this is likely to be a bit vague because of collaborators > etc..but an order of magnitude that we can all agree on would be great! > > And if anyone can estimate how many fish or fish lines even there are > then that would be fantastic! > > Many thanks, > > Caroline > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish From f.mueller from bham.ac.uk Thu May 21 12:14:57 2009 From: f.mueller from bham.ac.uk (Ferenc Mueller) Date: Thu May 21 12:18:32 2009 Subject: [Zbrafish] Re: Zbrafish Digest, Vol 48, Issue 8 In-Reply-To: <200905211703.n4LH38p12859@net.bio.net> References: <200905211703.n4LH38p12859@net.bio.net> Message-ID: HI Caroline, not a full answer to your question, but approx 200 in Europe and over 50 in the UK has been registered by EURMEDNET. My guess is around 7-800 word wide. It certainly was over 500 some 3-4 years ago. cheers ferenc -- Dr Ferenc M?ller Department of Medical and Molecular Genetics School of Clinical and Experimental Medicine College of Medical and Dental Sciences University of Birmingham, Institute of Biomedical Research B15 2TT, Edgbaston, Birmingham Tel: +44 121 414 2895 Fax: +44 121 414 2538 mobile: +44 7983420469 http://www.rch.bham.ac.uk/staff/Mueller.shtml From fritsa01 from gettysburg.edu Fri May 22 07:27:25 2009 From: fritsa01 from gettysburg.edu (FritzXC23) Date: Fri May 22 09:20:56 2009 Subject: [Zbrafish] Imaging Zebrafish Message-ID: <8f8dff4a-7965-485e-ad72-ea576199f16e@l32g2000vba.googlegroups.com> Hi All! My name is Sarah Fritz and I am undergraduate student from Gettysburg College. I am currently working on a research project that involves imaging fluorescent zebrafish hair cells. I am wondering if anyone has suggestions about the best way to get 5 dpf fish to lay on their sides without having to mount them. Thanks! Sarah Fritz From lhqkey from gmail.com Sun May 24 15:22:00 2009 From: lhqkey from gmail.com (Haiqiong) Date: Tue May 26 11:39:46 2009 Subject: [Zbrafish] Re: Imaging Zebrafish References: Message-ID: <5c58d5f5-1391-42d3-a550-d70a79644b91@l32g2000vba.googlegroups.com> Hi Sarah, Since it is fluorescent imaging, I guess you might be using some fluorescent dye for live embryos. Actually, it will be very hard to get good quality image without mounting the embryos, because it will be hard to go to high magnification without a coverslip. If you mean mounting the embryos but keeping them live for later use, you can use 0.5% agarose in E3 media with Tricaine. Haiqiong On May 22, 8:27?am, FritzXC23 wrote: > Hi All! > > My name is Sarah Fritz and I am undergraduate student from Gettysburg > College. I am currently working on a research project that involves > imaging fluorescent zebrafish hair cells. I am wondering if anyone has > suggestions about the best way to get 5 dpf fish to lay on their sides > without having to mount them. Thanks! > > Sarah Fritz From zhangch from ohsu.edu Sat May 23 14:20:43 2009 From: zhangch from ohsu.edu (Chao Zhang) Date: Tue May 26 11:40:25 2009 Subject: [Zbrafish] zebrafish inbred line References: <200905221703.n4MH3Pp16782@net.bio.net> Message-ID: Is zebrafish inbred line available for our research. AB, Tab or TU? Thanks. -------------- next part -------------- A non-text attachment was scrubbed... Name: not available Type: application/ms-tnef Size: 2314 bytes Desc: not available Url : http://www.bio.net/bionet/mm/zbrafish/attachments/20090523/6261d3cb/attachment.bin From Christian.Broesamle from med.uni-muenchen.de Tue May 26 13:49:25 2009 From: Christian.Broesamle from med.uni-muenchen.de (Christian Broesamle) Date: Tue May 26 13:55:39 2009 Subject: [Zbrafish] Re: Imaging Zebrafish In-Reply-To: <5c58d5f5-1391-42d3-a550-d70a79644b91@l32g2000vba.googlegroups.com> References: <5c58d5f5-1391-42d3-a550-d70a79644b91@l32g2000vba.googlegroups.com> Message-ID: Sarah, I imagine that you want to image quickly and possibly large numbers of larvae (screen?), since you don't want to mount them. 5 dpf ist difficult, because by this time they have inflated swim bladders and won't lie on their sides when anesthetized. Instead they may float belly up. At the last ZF Meeting there was a company (forgot the name but could find out) presenting a 96 well plate that had in-build little prisms that allowed imaging from below and getting a side view of the larvae. The draw-back was that this results in a rather long working distance and max. possible magnification they could get was with a 10x objective, if I remember correctly. I think it was pretty expensive too, but I see now reason why the plates couldn't be reused. Christian >Hi Sarah, > >Since it is fluorescent imaging, I guess you might be using some >fluorescent dye for live embryos. Actually, it will be very hard to >get good quality image without mounting the embryos, because it will >be hard to go to high magnification without a coverslip. If you mean >mounting the embryos but keeping them live for later use, you can use >0.5% agarose in E3 media with Tricaine. > >Haiqiong > > >On May 22, 8:27 am, FritzXC23 wrote: >> Hi All! >> >> My name is Sarah Fritz and I am undergraduate student from Gettysburg >> College. I am currently working on a research project that involves >> imaging fluorescent zebrafish hair cells. I am wondering if anyone has >> suggestions about the best way to get 5 dpf fish to lay on their sides >> without having to mount them. Thanks! >> >> Sarah Fritz > > >_______________________________________________ >Zbrafish mailing list >Zbrafish@net.bio.net >http://www.bio.net/biomail/listinfo/zbrafish -- *********************************** Christian Br?samle Department of Biochemistry Ludwig-Maximilians-Universit?t M?nchen Schillerstrasse 44 80336 M?nchen Germany tel: +49 89 2180 75-451 fax: +49 89 2180 75-415 *********************************** From Caroline.Parkin from sheffield.ac.uk Wed May 27 03:11:54 2009 From: Caroline.Parkin from sheffield.ac.uk (C Parkin) Date: Wed May 27 11:14:47 2009 Subject: [Zbrafish] Re Imaging Zebrafish In-Reply-To: <200905261703.n4QH3Ap12222@net.bio.net> References: <200905261703.n4QH3Ap12222@net.bio.net> Message-ID: <1243411914.4a1cf5ca2b330@webmail.shef.ac.uk> Hi Sarah, I also think the best way to image live fish is to put them in agarose - but remember to use low melting point agarose - or you'll fry your fish! I tend to melt a batch and put it in microtubes and keep it at 50C ready for use (either hotblock or waterbath). Don't add tricaine till you need it! I find mounting fish when the agarose is ready is very quick. Then it then depends if you have an upright or inverted scope. For inverted I like to use glass bottom mini petri dishes. For upright you can do the same and turn it upside down or use a water immersion lens. If you want to image lots of embryos on their side you could try a sideview plate (google it) or a optically clear 96 well plate and spin them down at low speed on a centrifuge - the majority should be laying on their side (all dependant on age though - works best for 2-4 days old embryos, or older if the swim bladder isn't inflated). This is only really suitable for an inverted scope. It is possible to use a silicone spacer (to make a small well) and put the embryo in the well (in E3) and put a cover slip on top. But your fish might move more and can't be left for a long time like this. I hope that's helpful, Caroline > > > On May 22, 8:27?am, FritzXC23 wrote: > > Hi All! > > > > My name is Sarah Fritz and I am undergraduate student from Gettysburg > > College. I am currently working on a research project that involves > > imaging fluorescent zebrafish hair cells. I am wondering if anyone has > > suggestions about the best way to get 5 dpf fish to lay on their sides > > without having to mount them. Thanks! > > > > Sarah Fritz > > > > > ------------------------------ > > Message: 2 > Date: Sat, 23 May 2009 12:20:43 -0700 > From: "Chao Zhang" > Subject: [Zbrafish] zebrafish inbred line > To: > Message-ID: > Content-Type: text/plain; charset="gb2312" > > Is zebrafish inbred line available for our research. > AB, Tab or TU? > Thanks. > -------------- next part -------------- > A non-text attachment was scrubbed... > Name: not available > Type: application/ms-tnef > Size: 2314 bytes > Desc: not available > Url : > http://www.bio.net/bionet/mm/zbrafish/attachments/20090523/6261d3cb/attachment-0001.bin > > ------------------------------ > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > > End of Zbrafish Digest, Vol 48, Issue 10 > **************************************** > From jalister from vcu.edu Wed May 27 12:17:08 2009 From: jalister from vcu.edu (James Lister) Date: Wed May 27 12:18:52 2009 Subject: [Zbrafish] Re Imaging Zebrafish In-Reply-To: <1243411914.4a1cf5ca2b330@webmail.shef.ac.uk> References: <200905261703.n4QH3Ap12222@net.bio.net> <1243411914.4a1cf5ca2b330@webmail.shef.ac.uk> Message-ID: <577D5DA784F848DB9DF6C74215FB847A@HGPCD4TSZ41> Here is a link to the sideview plate that Christian and Caroline mentioned... http://www.sideviewmicroplate.com/ -jim > -----Original Message----- > From: zbrafish-bounces@oat.bio.indiana.edu > [mailto:zbrafish-bounces@oat.bio.indiana.edu] On Behalf Of C Parkin > Sent: Wednesday, May 27, 2009 4:12 AM > To: zbrafish@oat.bio.indiana.edu > Subject: [Zbrafish] Re Imaging Zebrafish > > Hi Sarah, > > I also think the best way to image live fish is to put them > in agarose - but > remember to use low melting point agarose - or you'll fry > your fish! I tend to > melt a batch and put it in microtubes and keep it at 50C > ready for use (either > hotblock or waterbath). Don't add tricaine till you need it! > I find mounting > fish when the agarose is ready is very quick. > Then it then depends if you have an upright or inverted > scope. For inverted I > like to use glass bottom mini petri dishes. For upright you > can do the same and > turn it upside down or use a water immersion lens. > If you want to image lots of embryos on their side you could > try a sideview > plate (google it) or a optically clear 96 well plate and spin > them down at low > speed on a centrifuge - the majority should be laying on > their side (all > dependant on age though - works best for 2-4 days old > embryos, or older if the > swim bladder isn't inflated). This is only really suitable > for an inverted > scope. > It is possible to use a silicone spacer (to make a small > well) and put the > embryo in the well (in E3) and put a cover slip on top. But > your fish might > move more and can't be left for a long time like this. > > I hope that's helpful, > Caroline > > > > > > > > On May 22, 8:27?am, FritzXC23 wrote: > > > Hi All! > > > > > > My name is Sarah Fritz and I am undergraduate student > from Gettysburg > > > College. I am currently working on a research project > that involves > > > imaging fluorescent zebrafish hair cells. I am wondering > if anyone has > > > suggestions about the best way to get 5 dpf fish to lay > on their sides > > > without having to mount them. Thanks! > > > > > > Sarah Fritz > > > > > > > > > > ------------------------------ > > > > Message: 2 > > Date: Sat, 23 May 2009 12:20:43 -0700 > > From: "Chao Zhang" > > Subject: [Zbrafish] zebrafish inbred line > > To: > > Message-ID: > > > Content-Type: text/plain; charset="gb2312" > > > > Is zebrafish inbred line available for our research. > > AB, Tab or TU? > > Thanks. > > -------------- next part -------------- > > A non-text attachment was scrubbed... > > Name: not available > > Type: application/ms-tnef > > Size: 2314 bytes > > Desc: not available > > Url : > > > http://www.bio.net/bionet/mm/zbrafish/attachments/20090523/626 > 1d3cb/attachment-0001.bin > > > > ------------------------------ > > > > _______________________________________________ > > Zbrafish mailing list > > Zbrafish@net.bio.net > > http://www.bio.net/biomail/listinfo/zbrafish > > > > End of Zbrafish Digest, Vol 48, Issue 10 > > **************************************** > > > > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > From caroline.parkin from sheffield.ac.uk Thu May 28 03:55:20 2009 From: caroline.parkin from sheffield.ac.uk (Caroline Parkin) Date: Thu May 28 11:18:33 2009 Subject: [Zbrafish] Re:Number of ZF labs In-Reply-To: <200905261703.n4QH3Ap12222@net.bio.net> References: <200905261703.n4QH3Ap12222@net.bio.net> Message-ID: Hi all, Thanks everyone for their replies. Very helpful - cheers! I thought you might all be interested in ZFIN's response to my request for ZF researcher numbers: ZFIN currently contains: 589 labs 4821 registered users 4149 mutants affecting 2984 genes 87 deficiencies affecting 63 genes 26 translocations affecting 26 genes Thanks to Jonathan for those! Best wishes, Caroline On 26 May 2009, at 18:03, zbrafish-request@oat.bio.indiana.edu wrote: > Send Zbrafish mailing list submissions to > zbrafish@net.bio.net > > To subscribe or unsubscribe via the World Wide Web, visit > http://www.bio.net/biomail/listinfo/zbrafish > or, via email, send a message with subject or body 'help' to > zbrafish-request@net.bio.net > > You can reach the person managing the list at > zbrafish-owner@net.bio.net > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Zbrafish digest..." > > > Today's Topics: > > 1. Re: Imaging Zebrafish (Haiqiong) > 2. zebrafish inbred line (Chao Zhang) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Sun, 24 May 2009 13:22:00 -0700 (PDT) > From: Haiqiong > Subject: [Zbrafish] Re: Imaging Zebrafish > To: bionet-organisms-zebrafish@moderators.isc.org > Message-ID: > <5c58d5f5-1391-42d3-a550-d70a79644b91@l32g2000vba.googlegroups.com> > Content-Type: text/plain; charset=ISO-8859-1 > > Hi Sarah, > > Since it is fluorescent imaging, I guess you might be using some > fluorescent dye for live embryos. Actually, it will be very hard to > get good quality image without mounting the embryos, because it will > be hard to go to high magnification without a coverslip. If you mean > mounting the embryos but keeping them live for later use, you can use > 0.5% agarose in E3 media with Tricaine. > > Haiqiong > > > On May 22, 8:27 am, FritzXC23 wrote: >> Hi All! >> >> My name is Sarah Fritz and I am undergraduate student from Gettysburg >> College. I am currently working on a research project that involves >> imaging fluorescent zebrafish hair cells. I am wondering if anyone >> has >> suggestions about the best way to get 5 dpf fish to lay on their >> sides >> without having to mount them. Thanks! >> >> Sarah Fritz > > > > > ------------------------------ > > Message: 2 > Date: Sat, 23 May 2009 12:20:43 -0700 > From: "Chao Zhang" > Subject: [Zbrafish] zebrafish inbred line > To: > Message-ID: > Content-Type: text/plain; charset="gb2312" > > Is zebrafish inbred line available for our research. > AB, Tab or TU? > Thanks. > -------------- next part -------------- > A non-text attachment was scrubbed... > Name: not available > Type: application/ms-tnef > Size: 2314 bytes > Desc: not available > Url : http://www.bio.net/bionet/mm/zbrafish/attachments/20090523/6261d3cb/attachment-0001.bin > > ------------------------------ > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish > > End of Zbrafish Digest, Vol 48, Issue 10 > **************************************** From roy from biostatus.com Fri May 29 06:17:42 2009 From: roy from biostatus.com (Roy Edward) Date: Fri May 29 12:23:30 2009 Subject: [Zbrafish] Re: Imaging Zebrafish References: Message-ID: <6ff5ff01-ad60-4c3e-b1b6-e5876effc969@v2g2000vbb.googlegroups.com> On May 22, 8:27?am, FritzXC23 wrote: > Hi All! > > My name is Sarah Fritz and I am undergraduate student from Gettysburg > College. I am currently working on a research project that involves > imaging fluorescent zebrafish hair cells. I am wondering if anyone has > suggestions about the best way to get 5 dpf fish to lay on their sides > without having to mount them. Thanks! > > Sarah Fritz Hi Sarah Excuse the commercial response but we have developed a technology that should help in this respect. CyGEL/CyGEL Sustain are thermoreversible hydrogels that are a liquid when cooled and gel when warmed above room temperature. These were initially developed to allow fluorescent imaging of live non-adherent cells, but in collaboration with a zebrafish lab have proved very useful for zebrafish embryos. For the live embryo work we have found CyGEL Sustain to be best. It is optically inert and clear. It accepts embryo medium (40-60X is best to avoid over-dilution of the gel) and MS-222 and analogues. We have added MS-222 at a 2X final conc. compared to that used to initially anaesthetize the embryos, creating a reservoir of anaesthetic for the period of the procedure. Embryos can be moved into position by melting the gel, moving them and re-setting the gel. It can be used on slides with coverslips or in microplate wells. This process can be used for imaging or for micro-injection. In all cases embryos MUST be de-chorionated. Embryos can be recovered, live, after an hour in the gel, requiring only a gentle wash to remove residual gel and time to allow them to re- awaken! CyGEL (a PBS-based equivalent) is more suitable for fixed embryo imaging when special medium is not required, and is provided with 40X PBS. A poster describing CyGEL Sustain's use for live embryo manipulation will be presented by collaborators at the forthcoming zebrafish conference in Rome. I have also collaborated with C elegans and parasite researcher teams. Again we were able successfully to image and recover live organisms. Full details on CyGEL Sustain can be found at our website. I can also provide a copy of a previous poster I presented at SBS on our CyGEL technology. Very best regards Roy Roy Edward roy@biostatus.com Biostatus Ltd. Reg. Office: 56 Charnwood Road, Shepshed LEICS. LE12 9NP UK Company No: 3079239. Registered in England and Wales. Tel: +44 (0)1509 558163 Fax +44 (0)1509 651061 enquiry@biostatus.com www.biostatus.com From usuaoy from gmail.com Thu May 28 09:02:10 2009 From: usuaoy from gmail.com (Usua) Date: Fri May 29 12:23:51 2009 Subject: [Zbrafish] Problems with DNAse Message-ID: <3a0ce4a3-af9c-4efd-9dd5-addb5739c1b5@y9g2000yqg.googlegroups.com> Dear all, I am working with zebrafish embryos and gene expression. I use trizol for the RNA extraction, and then when I use DNAse before the RT PCR, I lose all the RNA in my sample. does anyone use a DNAse step in their RNA preps? I would like to know if it is neccessary to use dnases or if I could play directly the PCR. I know that my preps theoretically should not yield DNA product, but i want to be sure that the RTPCR products I am recieving are from an RNA template and not from DNA. Thank you in advance . From rburdine from Princeton.EDU Fri May 29 12:59:23 2009 From: rburdine from Princeton.EDU (Burdine, Rebecca D) Date: Fri May 29 13:11:42 2009 Subject: [Zbrafish] Problems with DNAse In-Reply-To: <3a0ce4a3-af9c-4efd-9dd5-addb5739c1b5@y9g2000yqg.googlegroups.com> References: <3a0ce4a3-af9c-4efd-9dd5-addb5739c1b5@y9g2000yqg.googlegroups.com> Message-ID: <790DAC09623D0A47ACDB126E3C52D8B61DC523@MBCLUSTER.pu.win.princeton.edu> Hi, TRIZOL uses guanidine isothiocyanate and acid phenol to isolate RNA based on the original paper by Chomczynski, P. and Sacchi, N. Anal. Biochem 162, 156 (1987). The acid phenol step should partition the RNA away from the DNA so you shouldn't need to DNAse if your preps are clean and you avoided the interface when separating the aqueous solution from the phenol. However, you shouldn't lose your RNA when you use DNAse regardless. Are you sure it is an RNAse free DNAse? You may want to purchase a new vial and give that a try. 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 Usua > Sent: Thursday, May 28, 2009 10:02 AM > To: bionet-organisms-zebrafish@moderators.isc.org > Subject: [Zbrafish] Problems with DNAse > > Dear all, > > I am working with zebrafish embryos and gene expression. I use trizol > for the RNA extraction, and then when I use DNAse before the RT PCR, I > lose all the RNA in my sample. > > does anyone use a DNAse step in their RNA preps? > > I would like to know if it is neccessary to use dnases or if I could > play directly the PCR. I know that my preps theoretically should not > yield DNA product, but i want to be sure that the RTPCR products I am > recieving are from an RNA template and not from DNA. > > Thank you in advance > . > > _______________________________________________ > Zbrafish mailing list > Zbrafish@net.bio.net > http://www.bio.net/biomail/listinfo/zbrafish