From qinh from cshl.edu Thu Jan 3 16:30:48 2008 From: qinh from cshl.edu (Qin, Hongtao) Date: Fri Jan 4 08:04:28 2008 Subject: [Drosophila] Preserving Drosophila by Freezing? Message-ID: Greeting all, I noticed from the following link that detailed protocols for preserving Drosophila by freezing were made available to the fly community in 1993. http://www.bio.net/bionet/mm/dros/1993-April/000011.html However, it seems like preserving Drosophila by freezing is still not an option for keeping fly stocks in many labs. Does anyone know why? Is it because the protocols were not reproducible by other labs, the cost was too high or other problems? Is any lab still trying hard to develop a better protocol? Thanks, Hongtao Hongtao Qin Cold Spring Harbor Laboratory 1 Bungtown Road Cold Spring Harbor, NY 11724 Phone: 516-367-6826 From arajamoh from uwo.ca Fri Jan 4 12:28:31 2008 From: arajamoh from uwo.ca (Arun Rajamohan) Date: Fri Jan 4 12:40:31 2008 Subject: [Drosophila] Re: Dros Digest, Vol 33, Issue 1 In-Reply-To: <200801041706.m04H6KL04078@net.bio.net> References: <200801041706.m04H6KL04078@net.bio.net> Message-ID: An interesting question. -A On Jan 4, 2008, at 12:06 PM, dros-request@oat.bio.indiana.edu wrote: > Send Dros mailing list submissions to > dros@net.bio.net > > To subscribe or unsubscribe via the World Wide Web, visit > http://www.bio.net/biomail/listinfo/dros > or, via email, send a message with subject or body 'help' to > dros-request@net.bio.net > > You can reach the person managing the list at > dros-owner@net.bio.net > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Dros digest..." > > > Today's Topics: > > 1. Preserving Drosophila by Freezing? (Qin, Hongtao) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Thu, 3 Jan 2008 16:30:48 -0500 > From: "Qin, Hongtao" > Subject: [Drosophila] Preserving Drosophila by Freezing? > To: > Message-ID: > > Content-Type: text/plain; charset="iso-8859-1" > > Greeting all, > > I noticed from the following link that detailed protocols for > preserving Drosophila by freezing were made available to the fly > community in 1993. > http://www.bio.net/bionet/mm/dros/1993-April/000011.html > However, it seems like preserving Drosophila by freezing is still > not an option for keeping fly stocks in many labs. Does anyone know > why? Is it because the protocols were not reproducible by other > labs, the cost was too high or other problems? Is any lab still > trying hard to develop a better protocol? > > Thanks, > Hongtao > > Hongtao Qin > Cold Spring Harbor Laboratory > 1 Bungtown Road > Cold Spring Harbor, NY 11724 > Phone: 516-367-6826 > > > > ------------------------------ > > _______________________________________________ > Dros mailing list > Dros@net.bio.net > http://www.bio.net/biomail/listinfo/dros > > End of Dros Digest, Vol 33, Issue 1 > *********************************** From arajamoh from uwo.ca Sat Jan 5 12:56:04 2008 From: arajamoh from uwo.ca (Arun Rajamohan) Date: Sat Jan 5 15:09:47 2008 Subject: [Drosophila] Re: Dros Digest, Vol 33, Issue 1 In-Reply-To: <200801041706.m04H6KL04078@net.bio.net> References: <200801041706.m04H6KL04078@net.bio.net> Message-ID: Hi All, Today I noticed that my previous mail contained only the first few words of what I intended to send. Sorry about that and here is the full version. ~ An interesting question. And I have also wondered about the same. My feeling is that some of the laboratories that attempted to reproduce the results published as early as 1990 did not continue their work due to a small learning curve associated with the technique. Also high variability in viability that will show up even with extremely minor change of conditions in the procedure - primarily embryonic stage selection for the procedure. I would be really interested in hearing from any other laboratories that have attempted cryopreserving D. melanogaster embryos and the various pitfalls that they encountered while trying to do it. For that matter I am also keen in knowing all the hearsay in this regard. -Arun On Jan 4, 2008, at 12:06 PM, dros-request@oat.bio.indiana.edu wrote: > Send Dros mailing list submissions to > dros@net.bio.net > > To subscribe or unsubscribe via the World Wide Web, visit > http://www.bio.net/biomail/listinfo/dros > or, via email, send a message with subject or body 'help' to > dros-request@net.bio.net > > You can reach the person managing the list at > dros-owner@net.bio.net > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Dros digest..." > > > Today's Topics: > > 1. Preserving Drosophila by Freezing? (Qin, Hongtao) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Thu, 3 Jan 2008 16:30:48 -0500 > From: "Qin, Hongtao" > Subject: [Drosophila] Preserving Drosophila by Freezing? > To: > Message-ID: > > Content-Type: text/plain; charset="iso-8859-1" > > Greeting all, > > I noticed from the following link that detailed protocols for > preserving Drosophila by freezing were made available to the fly > community in 1993. > http://www.bio.net/bionet/mm/dros/1993-April/000011.html > However, it seems like preserving Drosophila by freezing is still > not an option for keeping fly stocks in many labs. Does anyone know > why? Is it because the protocols were not reproducible by other > labs, the cost was too high or other problems? Is any lab still > trying hard to develop a better protocol? > > Thanks, > Hongtao > > Hongtao Qin > Cold Spring Harbor Laboratory > 1 Bungtown Road > Cold Spring Harbor, NY 11724 > Phone: 516-367-6826 > > > > ------------------------------ > > _______________________________________________ > Dros mailing list > Dros@net.bio.net > http://www.bio.net/biomail/listinfo/dros > > End of Dros Digest, Vol 33, Issue 1 > *********************************** From yehuditmi from gmail.com Sun Jan 6 08:27:00 2008 From: yehuditmi from gmail.com (yehudit) Date: Sun Jan 6 13:58:42 2008 Subject: [Drosophila] Drosophila- In situ hybridization problems Message-ID: <14648451.post@talk.nabble.com> Our lab has been doing whole mount in-situ hybridizations on Drosophila embryos using random-primed or PCR-produced DNA DIG probes for years. In the last year, we?ve had serious problems with high background, and some problems with weak signal. After examing all components, we see that the most significant factor is the enzyme used for PCR! Most suprisingly, using Polymerase from Roche gives us background problems that are often the worst, and better results (but not consistently good) with enzymes like Super-Nova polymerase from NutriCyte (Buffalo, NY) and others. This is all in a system in which we?re: using Roche anti-DIG; pre-absorbing the Antibody with embryos; and blocking by methods that previously worked well. The background problems are not sequence specific, and arise with all probes. Does anyone have suggestions about this problem? Or any general suggestions for improved ways of reducing background? -- View this message in context: http://www.nabble.com/Drosophila--In-situ-hybridization-problems-tp14648451p14648451.html Sent from the Bio.net - Dros mailing list archive at Nabble.com. From marko from radulovic.net Tue Jan 8 08:22:46 2008 From: marko from radulovic.net (Marko Radulovic) Date: Tue Jan 8 13:59:30 2008 Subject: [Drosophila] S2 Cell growth vs temperature Message-ID: <000001c851f9$8fb99570$af2cc050$@net> Dear Robert, I read your post at bionet, unfortunately I can not help you as I do not have much experience with S2 cells. As I see that you grow S2 cells routinely, I wanted to ask you for a favor to tell me how you do it. I need to grow large amounts of these cells, which is easiest done in suspension, but my experience is that they do not grow well in suspension, although they should. I have varied the speed of mixing etc, but no improvement. I used big glass erlenmayer flasks, then tried big plastic rotating bottles, but little success. I use Schneiders medium with 10%FCS plus glutamine/pen/strep. I would very much appreciate if you could find time to tell me about the conditions in which S2 cells would grow well in suspension. Many thanks Marko From sa08383 from wotan.mdacc.tmc.edu Tue Jan 8 17:38:00 2008 From: sa08383 from wotan.mdacc.tmc.edu (Kathleen Gajewski) Date: Tue Jan 8 18:10:08 2008 Subject: [Drosophila] Drosophila- In situ hybridization problems In-Reply-To: <14648451.post@talk.nabble.com> References: <14648451.post@talk.nabble.com> Message-ID: <5749e2f631705d5363573bedab9975c7@wotan.mdacc.tmc.edu> A number of years back I had problems with staining that were caused by using Roche anti-DIG Ab that was purchased separately (as opposed to coming as part of a transcription kit). I don't know it that's still a problem, but it may be worth looking into. KG On Jan 6, 2008, at 7:27 AM, yehudit wrote: > > Our lab has been doing whole mount in-situ hybridizations on Drosophila > embryos using random-primed or PCR-produced DNA DIG probes for years. > In the > last year, we’ve had serious problems with high background, and some > problems with weak signal. After examing all components, we see that > the > most significant factor is the enzyme used for PCR! Most suprisingly, > using > Polymerase from Roche gives us background problems that are often the > worst, > and better results (but not consistently good) with enzymes like > Super-Nova > polymerase from NutriCyte (Buffalo, NY) and others. This is all in a > system > in which we’re: using Roche anti-DIG; pre-absorbing the Antibody with > embryos; and blocking by methods that previously worked well. The > background > problems are not sequence specific, and arise with all probes. > Does anyone have suggestions about this problem? Or any general > suggestions > for improved ways of reducing background? > > -- > View this message in context: > http://www.nabble.com/Drosophila--In-situ-hybridization-problems- > tp14648451p14648451.html > Sent from the Bio.net - Dros mailing list archive at Nabble.com. > > > _______________________________________________ > Dros mailing list > Dros@net.bio.net > http://www.bio.net/biomail/listinfo/dros From johncumbers from gmail.com Thu Jan 10 15:27:46 2008 From: johncumbers from gmail.com (John Cumbers) Date: Thu Jan 10 16:08:26 2008 Subject: [Drosophila] How many cells in a Drosophila head? Message-ID: Hi all, I'm trying to estimate how many cells are in a Drosophila head. I realise this is a difficult question and would depend on environment and nutrition, but a ball park figure with some references would be great. Many thanks, John -- John Cumbers, Graduate Student Biology and Medicine Brown University, Box G-W Providence, Rhode Island, 02912, USA Tel USA: +1 401 523 8190, Fax: +1 401 863-2166 UK to USA: 0207 617 7824 From sciencebook from mac.com Thu Jan 10 23:11:25 2008 From: sciencebook from mac.com (sciencebook@mac.com) Date: Fri Jan 11 09:20:49 2008 Subject: [Drosophila] Re: Mouth Pieces References: Message-ID: You can use a sterile blue tip. Clip off the point so you can suck in the right amount of air. From kbergman from keene.edu Fri Jan 11 12:03:17 2008 From: kbergman from keene.edu (Bergman, Ken) Date: Fri Jan 11 14:14:13 2008 Subject: [Drosophila] sources of CO2 stages Message-ID: <3E58EA5EABA507428E70A48D95477008315306@washington.keene.edu> All, dros@net.bio.net For years I've used the rectangular flat white box CO2 stages manufactured by the Harvard BioLabs Machine Shop, which went out of business a year or two ago. I've found the names of two other suppliers of CO2 staging (Genesee and LabScientific) but don't know much about their products and services. Can any share information about CO2 stages from those supplies or advise me about other reliable suppliers or sources? I'll check the Dros.net archives for replies, as I'm not currently subscribed, or you can send messages to kbergman@keene.edu. Thanks very much for your help. Ken Bergman Prof. of Biology Keene State College Keene, NH 03435 From nellygidas from yahoo.fr Fri Jan 11 10:54:27 2008 From: nellygidas from yahoo.fr (Nelly) Date: Fri Jan 11 14:15:40 2008 Subject: [Drosophila] question: mortality problems in drosophila Message-ID: <225538.20356.qm@web26012.mail.ukl.yahoo.com> Dear all, I?d like to call for advice or suggestions from people of the Drosophila community who have encountered mortality issues in Drosophila lines. Six months ago, the Drosophila pseudoobscura populations raised in my lab have suddenly shown high levels of mortality (80-90%), when the flies are 5-6 day old, particularly males. The flies that are still alive in the bottles look normal and have subsequent normal survival, although they seem to be a bit less productive. The problem has persisted despite trying several different procedures: 1) The room where the flies are handled was thoroughly cleaned (with dilute bleach and 90% ethanol) several days in succession and much of our equipment (plasticware, yeast etc.) was replaced. 2) An experiment was carried out to determine if the food was hosting some microorganism that could be the cause of the infection. This involved raising males and females separately on 3 different types of food medium; normal (cornmeal molasses medium), sugar agar or a potato based medium. Flies were transferred to fresh vials of the appropriate medium every day for around 10-14 days. The vials were then observed to see if anything grew on these substrates; it was assumed we could detect any microorganism present in the food. There was no obvious presence of a microorganism growing on any substrate and the survival of the flies did not differ according to which substrate they were kept on. 3) Finally, an antibiotic treatment (0.025% tetracycline) was introduced to the food. Flies cultured on this substrate were less productive and pupated faster than those on normal food. The offspring of the treated generation survived normally for one generation. Unfortunately, the following generation again displayed the same symptoms of mass mortality at 5 days old. I have contacted companies that can identify microorganisms and suggest treatments but they were not able to respond our queries. I would be really grateful if anyone has any suggestions on other procedures that I could undertake or who to contact who would be able to suggest solutions to determine the problem. Thanks! Nelly Gidaszewski Research associate Department of Animal and Plant Sciences University of Sheffield UK email: n.gidaszewski@sheffield.ac.uk _____________________________________________________________________________ Ne gardez plus qu'une seule adresse mail ! Copiez vos mails vers Yahoo! Mail http://mail.yahoo.fr From hotbacteria from rediffmail.com Fri Jan 11 14:42:55 2008 From: hotbacteria from rediffmail.com (tarun gupta) Date: Fri Jan 11 15:46:29 2008 Subject: [Drosophila] Re: How many cells in a Drosophila head? Message-ID: <20080111194255.22061.qmail@f5mail-237-204.rediffmail.com> ? Hi John, Although I am not an expert in Drosophla Biology but perhaps you can find it experimentally by FACS analysis. I am sure you can get a fairly accurate figure. You may also want to do that for various developmental Stages. Kind Regards, Tarun Gupta MSc Human Genomics National Centre for Human Genome Studies and Research Panjab University Chandigarh-India Mob: +91-9888237906 Appeal: Let's not make information inaccessible to masses; Let's publish in Open Access Journals! ****************************** ******************* ~Exciting Science forum: http://groups.google.com/group/exciting-science ~About Me: http://hotbacteria.blogspot.com ~Scientific Information Sharing Resource: http://sisr.blogspot.com ~My Department: http://nchgsr.puchd.ac.in ************************************************* > John wrote: >Hi all, >I'm trying to estimate how many cells are in a Drosophila head. I realise >this is a difficult question and would depend on environment and nutrition, >but a ball park figure with some references would be great. >Many thanks, >John > > >-- >John Cumbers, Graduate Student >Biology and Medicine >Brown University, Box G-W >Providence, Rhode Island, 02912, USA >Tel USA: +1 401 523 8190, Fax: +1 401 863-2166 >UK to USA: 0207 617 7824 From mmspring from ncsu.edu Mon Jan 14 22:27:41 2008 From: mmspring from ncsu.edu (mmspring@ncsu.edu) Date: Tue Jan 15 09:27:53 2008 Subject: [Drosophila] Dissecting drosophila fat bodies and nile red staining Message-ID: <41066.69.134.151.59.1200367661.squirrel@webmail.ncsu.edu> Hi All, I am wondering if anyone has an efficient method for dissecting and isolating drosophila fat bodies from 3rd instar larvae so that they can easily be stained. Also I am using Nile Red as the stain to visualize fat bodies and was wondering if anyone else had used Nile Red with a regular compound fluorescent microscope. Most of the literature I have found describes using a confocal microscope and I am looking for the best buffers to use and the right dilution of stain when making my slides. Thanks for any help I can get. Mastafa Springston Research Assistant Greg Gibson Laboratory North Carolina State University Raleigh, NC USA From runahamid from googlemail.com Wed Jan 16 08:13:24 2008 From: runahamid from googlemail.com (Runa Hamid) Date: Thu Jan 17 08:29:27 2008 Subject: [Drosophila] Viability test for drosophila tissue Message-ID: <2e1863ea0801160513mae801f2y37b4d6f091e20d89@mail.gmail.com> Hi all, I am doing live imaging in drosophila III instar larval brain. During imaging the brain is kept in a specific buffer. The brain is expressing GFP tagged protein in all cholinergic neurons.I need to show experimentally that the brain is alive for the time I require for the imaging (20-30 min). Can anyone suggest a Viability test for "TISSUE" (more specifically drosophila larval brain) where I can determine the degree of cell death in the tissue. I will be grateful if one of you could help me with the suggestions or protocol. Thanks in advance, Runa Department of Biological Sciences Tata Institute of Fundamental Research Colaba, Mumabai 400 005, India. From artyw2 from yahoo.com Thu Jan 17 11:35:58 2008 From: artyw2 from yahoo.com (artyw2@yahoo.com) Date: Thu Jan 17 12:24:07 2008 Subject: [Drosophila] Re: Viability test for drosophila tissue References: Message-ID: <1e84621a-3820-4638-9de7-5b12f5539039@e6g2000prf.googlegroups.com> On Jan 16, 8:13 am, "Runa Hamid" wrote: > Hi all, > > I am doing live imaging in drosophila III instar larval brain. During > imaging the brain is kept in a specific buffer. The brain is expressing GFP > tagged protein in all cholinergic neurons.I need to show experimentally > that the brain is alive for the time I require for the imaging (20-30 min). > Can anyone suggest a Viability test for "TISSUE" (more specifically > drosophila larval brain) where I can determine the degree of cell death in > the tissue. > I will be grateful if one of you could help me with the suggestions or > protocol. > > Thanks in advance, Larval brains label very well with 35S methionine. You might also use cell death stains such as acridine orange, although I do not know whether cell death will be complete enough during such a short interval to stain well. From dmerrill from ksu.edu Thu Jan 17 15:02:59 2008 From: dmerrill from ksu.edu (Arthropod Genomics) Date: Thu Jan 17 16:02:46 2008 Subject: [Drosophila] Registration open for Arthropod Genomics Symposium in Kansas City Message-ID: <001101c85943$f5901300$78738281@ECOGENOFFICE> SYMPOSIUM: NEW INSIGHTS FROM ARTHROPOD GENOMES April 11-13, 2008, in Kansas City Registration is now open to attend the 2nd Annual Arthropod Genomics Symposium, April 11 - 13, 2008, in Kansas City. A brochure and complete information with links for registration and hotel reservations can be downloaded at www.ksu.edu/agc/symposium.shtml. SYMPOSIUM PROGRAM: The main symposium sessions will take place on Friday-Saturday, April 11-12. Speakers will present new insights from genomic approaches in arthropods and describe the development of tools for genomic analysis. Optional workshops are scheduled for Thursday and Friday evenings. An evening of jazz and KC barbeque is planned for Saturday night. On Sunday morning, participate in a roundtable discussion with the ArthropodBase Consortium. Activities will conclude by noon on Sunday. FEATURED SPEAKERS: John Kenneth Colbourne, Indiana University, Preservation, expansion and invention of crustacean genes with reference to insect genomes. Christine G. Elsik, Georgetown University, Unusual base composition of the honey bee genome. Sarjeet S. Gill, University of California, Riverside, Mosquito midgut interactions with bacterial toxins. Catherine A. Hill, Purdue University, Tick genome organization and evolution. Thomas Kaufman, Indiana University, The latest news from CNN: What the 12 sequenced Drosophila genomes have told us about rapidly evolving genes and positive selection. J. Robert Manak, University of Iowa, Empirical annotation of arthropod genomes using tiled genomic microarrays. Subbaratnam Muthukrishnan, Kansas State University, Functional genomics of insect chitin metabolism. Hugh M. Robertson, University of Illinois at Urbana-Champaign, What we've learned about the insect chemoreceptors from arthropod genome projects. Bruce R. Schatz, University of Illinois at Urbana-Champaign, BeeSpace: Interactive functional analysis of arthropod genomic data. Jeff Stuart, Purdue University, Avirulence, sex determination, and a physical map of the Hessian fly genome. Judy Willis, University of Georgia, Insect cuticular proteins: Annotation, proteomics, expression, evolution. Evgeny Zdobnov, University of Geneva, Medical School, Switzerland, A comparative perspective on insect genomes. POSTER SESSIONS: There will be two poster sessions, limited to first 150 abstracts received before Friday, February 29. A few platform presentations will be chosen from submitted poster abstracts. WORKSHOPS AND ROUNDTABLE DISCUSSION: On Thursday evening, a workshop on "Community Contributions to Genome Annotation" will feature a presentation on use of the Apollo Genome Annotation Curation Tool by Dr. Chris Elsik (BeeBase). On Friday evening, Dave Clements (NESCent) and Scott Cain (CSHL) will lead a workshop, "Chado Databases and Integration with GMOD Tools." Throughout the meeting, arthropod genome database and bioinformatics tool developers will be available for individual training. On Sunday morning, participate in a roundtable discussion led by members of the ArthropodBase Consortium regarding the generation of integrated arthropod genome databases and tools for genome analysis, and community curation. Symposium attendees are invited to participate in these additional events. VENUE: The symposium will take place at the historic Muehlebach Hotel (operated by Marriott) in downtown Kansas City. KANSAS CITY JAZZ AND BARBEQUE: Participants are encouraged to stay Saturday night for an optional evening of jazz and KC barbeque in the historic 18th and Vine district. REGISTRATION: The registration fee will be $275 ($150 for graduate and undergraduate students), and will include a welcome reception Thursday evening, breakfast and lunch on Friday and Saturday, and light refreshments at the Friday poster session. Additional fees apply for the Apollo Workshop Thursday evening and Saturday night dinner. INFORMATION: Contact Doris Merrill at dmerrill@k-state.edu or 785-532-3482. To receive future Symposium announcements, send your contact information to dmerrill@k-state.edu. SYMPOSIUM WEBSITE: www.k-state.edu/agc/symposium.shtml DEADLINES: February 29, 2008 - Poster abstracts (limited to first 150 received) February 29, 2008 - Early registration March 20, 2008 - Hotel reservations Susan J. Brown, Professor Director, Center for Genomic Studies on Arthropods Affecting Human, Animal and Plant Health by Doris Merrill, Program Coordinator K-State Arthropod Genomics Center Division of Biology, Kansas State University 116 Ackert Hall, Manhattan, KS 66506-4901 (785) 532-3482, dmerrill@k-state.edu www.k-state.edu/agc From samarezzat0 from yahoo.com Fri Jan 25 08:55:00 2008 From: samarezzat0 from yahoo.com (Samar Ezzat) Date: Fri Jan 25 10:36:55 2008 Subject: [Drosophila] Drosophila embryo injection Message-ID: <54676.72295.qm@web34815.mail.mud.yahoo.com> Hallo everybody , I tried to inject Drosophila embryos using double -sided tape , i can dechorionate embryos manually and row them on other double -sided tape but when i put 10 s oil even in very small amount embryos slip . i will be very happy if i get any suggestion . Thanks a lot --------------------------------- Looking for last minute shopping deals? Find them fast with Yahoo! Search. From sa08383 from wotan.mdacc.tmc.edu Fri Jan 25 12:28:42 2008 From: sa08383 from wotan.mdacc.tmc.edu (Kathleen Gajewski) Date: Fri Jan 25 12:46:09 2008 Subject: [Drosophila] Drosophila embryo injection In-Reply-To: <54676.72295.qm@web34815.mail.mud.yahoo.com> References: <54676.72295.qm@web34815.mail.mud.yahoo.com> Message-ID: <6cfdd4b1a711e29f97be21e4362125e6@wotan.mdacc.tmc.edu> Exactly which brand of tape are you using? I use Scotch double stick style #MMM-665. If that's what you are using, then you may have to try another oil. I dechorionate with bleach, wash with lots of H2O, some 0.2% tritonX-100, and more H2O. I line the embryos up on a grape plate and press the slide with the tape on top of them, hard enough for them to stick, but not hard enough to smash them. I've had very good results with this method. Good luck KG On Jan 25, 2008, at 7:55 AM, Samar Ezzat wrote: > Hallo everybody , > > I tried to inject Drosophila embryos using double -sided tape , i > can dechorionate embryos manually and row them on other double -sided > tape but when i put 10 s oil even in very small amount embryos slip > . > > i will be very happy if i get any suggestion . > > Thanks a lot > > > --------------------------------- > Looking for last minute shopping deals? Find them fast with Yahoo! > Search. > _______________________________________________ > Dros mailing list > Dros@net.bio.net > http://www.bio.net/biomail/listinfo/dros From jaiganesha from gmail.com Sat Jan 26 13:52:36 2008 From: jaiganesha from gmail.com (Gurudatta Baraka) Date: Sun Jan 27 09:59:16 2008 Subject: [Drosophila] Re: Dros Digest, Vol 33, Issue 12 In-Reply-To: <200801261703.m0QH3eL06023@net.bio.net> References: <200801261703.m0QH3eL06023@net.bio.net> Message-ID: <84b2a75a0801261052l35caaf39o80c2a362c4ed5bf2@mail.gmail.com> hi nice to see the details of the glue tape usally peopl use halocarobon 700 for the injections do u use any specific oil with the scorch tape ? pelase suggest the product catalogue number etc thanks ' datta., On Jan 26, 2008 12:03 PM, wrote: > Send Dros mailing list submissions to > dros@net.bio.net > > To subscribe or unsubscribe via the World Wide Web, visit > http://www.bio.net/biomail/listinfo/dros > or, via email, send a message with subject or body 'help' to > dros-request@net.bio.net > > You can reach the person managing the list at > dros-owner@net.bio.net > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Dros digest..." > > > Today's Topics: > > 1. Re: Drosophila embryo injection (Kathleen Gajewski) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Fri, 25 Jan 2008 11:28:42 -0600 > From: Kathleen Gajewski > Subject: Re: [Drosophila] Drosophila embryo injection > To: Samar Ezzat > Cc: dros@magpie.bio.indiana.edu > Message-ID: <6cfdd4b1a711e29f97be21e4362125e6@wotan.mdacc.tmc.edu> > Content-Type: text/plain; charset=US-ASCII; format=flowed > > Exactly which brand of tape are you using? I use Scotch double stick > style #MMM-665. If that's what you are using, then you may have to try > another oil. > > I dechorionate with bleach, wash with lots of H2O, some 0.2% > tritonX-100, and more H2O. I line the embryos up on a grape plate and > press the slide with the tape on top of them, hard enough for them to > stick, but not hard enough to smash them. I've had very good results > with this method. > > Good luck > > KG > > > > > > On Jan 25, 2008, at 7:55 AM, Samar Ezzat wrote: > > > Hallo everybody , > > > > I tried to inject Drosophila embryos using double -sided tape , i > > can dechorionate embryos manually and row them on other double -sided > > tape but when i put 10 s oil even in very small amount embryos slip > > . > > > > i will be very happy if i get any suggestion . > > > > Thanks a lot > > > > > > --------------------------------- > > Looking for last minute shopping deals? Find them fast with Yahoo! > > Search. > > _______________________________________________ > > Dros mailing list > > Dros@net.bio.net > > http://www.bio.net/biomail/listinfo/dros > > > ------------------------------ > > _______________________________________________ > Dros mailing list > Dros@net.bio.net > http://www.bio.net/biomail/listinfo/dros > > End of Dros Digest, Vol 33, Issue 12 > > From hancockdh from Cardiff.ac.uk Thu Jan 31 08:38:51 2008 From: hancockdh from Cardiff.ac.uk (Daniel Hancock) Date: Thu Jan 31 08:54:59 2008 Subject: [Drosophila] Antibody stain variation (Dan Hancock) Message-ID: <47A1CF6B0200000F001039D8@zgrw01.cf.ac.uk> Dear All, When we have performed antibody stainings on fixed Drosophila embryos we have noticed that there is significant variation in intensity of staining for embryos of the same developmental stage. The embryos were all fixed together, in the same fix (i.e not a staining of multiple fixed embryos pooled together), and fixed using either paraformaldehyde or formaldehyde. This occurs with various antibodies. As well as variation between embryos we sometimes see variation in intensity of staining within the same embryo. If anyone has any ideas on why such variation is occurring and what we can do to reduce it then your advice would be appreciated. Many Thanks, Dan Hancock Cardiff University ------ Dan HANCOCK School of Biosciences Cardiff University Main Building Park Place Cardiff, CF10 3TL Wales, UK Tel: +44 (0) 29 2087 5884 Email: hancockdh@cf.ac.uk From stephanie_mohr from hms.harvard.edu Thu Jan 31 14:36:35 2008 From: stephanie_mohr from hms.harvard.edu (Stephanie Mohr) Date: Thu Jan 31 15:11:19 2008 Subject: [Drosophila] Assay Development Position at DRSC Message-ID: <47A22343.8010402@hms.harvard.edu> The Drosophila RNAi Screening Center (DRSC) at Harvard Medical School is seeking a motivated scientist with Drosophila experience to do independent research in this community-oriented center. Research focus on RNAi knockdown and over-expression of non-coding RNAs plus construction of new screening reagents (e.g. for over-expression, new cell lines), assay development and assay optimization. Opportunity to present and publish original research. Will be encouraged to collaborate with others. Access to specialized reagents and equipment, including for state-of-the-art high-content imaging. Excellent opportunity for a PhD interested in hands-on benchwork but seeking a non-traditional postdoc. Harvard is an award-winning employer with excellent benefits. Please send cover letter and CV to lmancini@receptor.med.harvard.edu with "DRSC position" in subject line.