DROSOPHILA INFORMATION NEWSLETTER Volume 13, January 1994 The Drosophila Information Newsletter has been established with the hope of providing a timely forum for informal communication among Drosophila workers. The Newsletter will be published quarterly and distributed electronically, free of charge. We will try to strike a balance between maximizing the useful information included and keeping the format short; priority will be given to genetic and technical information. Brevity is essential. If a more lengthy communication is felt to be of value, the material should be summarized and an address made available for interested individuals to request more information. Submitted material will be edited for brevity and arranged into each issue. Research reports, lengthy items that cannot be effectively summarized, and material that requires illustration for clarity should be sent directly to Jim Thompson (THOMPSON@AARDVARK.UCS.UOKNOR.EDU) for publication in DIS. Materials appearing in the Newsletter will be reprinted in DIS. Back issues of DIN are available from FlyBase in the directory flybase/news or in News/ when accessing FlyBase with Gopher. Material appearing in the Newsletter may be cited unless specifically noted otherwise. Material for publication should be submitted by e-mail. Figures and photographs cannot be accepted at present. Send technical notes to Carl Thummel and all other material to Kathy Matthews. The e-mail format does not allow special characters to be included in the text. Both superscripts and subscripts have been enclosed in square brackets; the difference should be obvious by context. Bold face, italics, underlining, etc. cannot be retained. Please keep this in mind when preparing submissions. To maintain the original format when printing DIN, use Courier 10cpi font on a standard 8.5" x 11" page with 1" margins. Drosophila Information Newsletter is a trial effort that will only succeed if a broad segment of the community participates. If you have information that would be useful to your colleagues, please take the time to pass it along. The editors: Carl Thummel Kathy Matthews Dept. of Human Genetics Dept. of Biology Eccles Institute - Bldg. 533 Indiana University University of Utah Bloomington, IN 47405 Salt Lake City, UT 84112 812-855-5782; FAX/2577 801-581-2937; FAX/5374 MATTHEWK@INDIANA.EDU CTHUMMEL@HMBGMAIL.MED.UTAH.EDU MATTHEWK@INDIANA.BITNET *** To add your name to the Newsletter distribution list, send one of the following E-mail messages from the account at which you wish to receive DIN. Via Internet -- To: LISTSERV@IUBVM.UCS.INDIANA.EDU Subject: Message: SUB DIS-L Your real name Via Bitnet -- To: LISTSERV@IUBVM Subject: Message: SUB DIS-L Your real name LISTSERV will extract your user name and node from the E-mail header and add you to the list. Use your Internet address if you have one. You will receive confirmation by E-mail if you have successfully signed on to the list. If you are on the list and do not wish to receive DIN, or you want to remove a soon-to- be-defunct address, replace SUB in the above message with UNS. The SUB command can also be used to correct spelling errors in your real name; the new entry will simply replace the old as long as it was sent from the same USERID@NODE address. *** DIN Vol. 13 TABLE OF CONTENTS >Introduction to Drosophila Information Newsletter >How to subscribe to the Newsletter >TABLE OF CONTENTS >ANNOUNCEMENTS >Crete Developmental Biology Workshop >1994 and 1995 US Drosophila Conferences >Bloomington Stock Center news >FlyBase and bionet.drosophila reminder >REQUESTS FOR MATERIALS >Wild-caught bb mutants >MATERIALS AVAILABLE >Monoclonal antibody against embryonic chordotonal organs >TECHNICAL NOTES >Preparation of DNA from single embryos for PCR *** ANNOUNCEMENTS CRETE DEVELOPMENTAL BIOLOGY MEETING An EMBO International Workshop on the MOLECULAR AND DEVELOPMENTAL BIOLOGY OF DROSOPHILA will be held at Kolymbari, Crete (Greece), June 19 - 25, 1994. The workshop will be co-sponsored by the U. of Crete and the Molecular Biology and Biotechnology Inst. of the Research Center of Crete. As has been the case in previous years, the objective of the workshop is to discuss topics in gene organization and expression, early development, pattern formation, developmental neurobiology and evolution. Approximately 80 participants will be selected by vote by the organizing committee from applicants. The participants are expected to contribute to the subject coverage. Applications should include a short summary of research interests and a brief C.V. if such information is thought to be important in facilitating the selection process. Please note that acceptance to the meeting is strictly limited to the individuals accepted. The deadline for applications is January 10, 1994. For logistical purposes we need to adhere strictly to this deadline. Applications should be sent to: CRETE WORKSHOP c/o Dr. S. Artavanis-Tsakonas Yale University School of Medicine - BCMM #236 295 Congress Avenue - P.O. Box 9812 New Haven, Connecticut 06536-0812 - U.S.A. A registration fee of $400.00 will be remitted upon acceptance. Local expenses in Crete will be covered. Participants are expected to finance their travel to Crete. However, a small number of grants in aid for partial travel support may become available. Preference will be given to junior investigators. The Organizing Committee: M. Ashburner (Cambridge U.), S. Artavanis-Tsakonas (Yale U.), B. Baker (Stanford U.), A. Bucheton, (CNRS), L. Cooley, (Yale U.), V. Corces (Johns Hopkins U.), M. Gatti (U. of Rome), W. Gehring (Basel U.), W. Gelbart (Harvard U.), D. Glover (Dundee U.), C. Goodman (USC - Berkeley), D. Hogness (Stanford U.), D. Ish-Horowicz (ICRF-U. of Oxford), H. Jackle (Max Planck - Gottingen), F. Kafatos (EMBL), D. Kankel (Yale U.), K. Louis (U. of Crete), P. Lawrence (MRC - Cambridge), J. Modolell, (U. Autonoma de Madrid), G. Morata (U. Autonoma de Madrid), G. Rubin (USC - Berkeley), R. Saint (U. of Adelaide), G. Schubiger (U. of Washington), B. Shilo (Weizmann Inst.), P. Simpson (Strasbourg), A. Spradling (Carnegie Inst. of Washington). *** 35th ANNUAL DROSOPHILA RESEARCH CONFERENCE The next US Drosophila Conference will be held in Chicago, Illinois, at the Sheraton Chicago Hotel, 301 East North Water St., April 20-24, 1994. The deadline for advance registration is February 14, 1994. Advance registration is $110 for GSA members ($55 for graduate students), and $140 for non-members ($75 for graduate students). The deadline for receipt of abstracts has passed. The Program Chairman is Victoria Finnerty, Biology Dept., Emory U., 1510 Clifton Rd., Atlanta, GA 30322, USA (e- mail: VICTORIA@BIOLOGY.EMORY.EDU). Contact The Genetics Society of America, 9650 Rockville Pike, Bethesda, MD 20814-3998 (301- 571-1825) for registration and housing forms. The 1995 US conference will be held April 5-9 in Atlanta, Georgia. *** BLOOMINGTON STOCK CENTER NEWS * USE STATISTICS FOR 1993 -- 16,785 stocks were shipped from the Bloomington Stock Center in 1993. This represents a 36% increase compared to 1992, and a 401% increase over the past 5 years. Weekly averages for the last quarter of 1993 were 58 requests for stocks or information, 48 shipments, and 442 stocks shipped. 35% of those shipments went outside the USA: 23% to European Community countries, 3.6% to Canada, 3.6% to Japan, 2.4% to Israel, and 2.9% to an assortment of other countries. Deficiency stocks continue to be the most requested category. * USER SURVEY FOR NIH -- At present NSF is the only funding agency providing support for Drosophila stock centers. NIH has expressed an interest in sharing support for the Bloomington center with NSF if we can demonstrate that a large proportion of our users are funded by NIH. We will be distributing user surveys over the next few weeks to gather information about our users' sources of research support. We apologize for one more piece of paperwork/e-mail (but imagine how we feel!), and will very much appreciate your prompt response. * SEND US YOUR REFERENCES -- In response to Vice-President Gore's good-government activities, NSF is asking stock centers funded by its program to provide documentation of specific scientific advances that were supported by materials from the center. We have started maintaining a database of publications that made significant use of stocks received from the Bloomington Stock Center. It would be extremely helpful if you would send us references for your relevant papers (now and in the future) with a VERY brief description of the role of center stocks, for example, 'used P insert at 25F to clone gene x'. Send e-mail to MATTHEWK@INDIANA.EDU. * HELP US COPE -- As you can see from the statistics cited above, use of the center has increased dramatically over the past five years. It is increasingly important that all of our users use the center responsibly. Please help us maintain our current level of service by complying with the following requests: 1. We are funded as a research resource, not as a teaching resource. Please do not order stocks from us for teaching purposes unless those genotypes are not available elsewhere, and PLEASE!!! do not refer teachers, parents, and science fair advisors to us for help with their student projects. 2. If the same stocks are available from both Bloomington and the Umea Stock Center, workers in European labs should order stocks from Umea. It is quite time-consuming for us to routinely check European requests against the Umea stock list and we will soon stop doing this altogether. Requests from European labs for non-P stocks must note that the requested stocks are not available from Umea or the order will be returned by post unfilled. 3. Use e-mail if you have it (to MATTHEWK@INDIANA.EDU). Sending a FAX is very time-consuming compared to responding to an e-mail message and FAXes usually have to be trimmed or folded to fit into our file folders. 4. When you place your first order from a new address explicitly state that the address provided is a new one. Mailing labels are automatically filled in from our 'address' database (1,760 names and addresses) and your stocks may go to your old address if you count on us to notice that you have moved. We often catch these, but it creates extra work to correct them after your request has been entered into the database. Also, it is helpful if you always use the same form of your name when ordering. It would be very helpful if some of you Johnsons and Martins would change your last names (just kidding on that one). 5. Order efficiently. Whenever possible, order by stock number; check your request for typos in the stock numbers before sending it. If you do not have access to our stock list, include the gene symbol in your request (e.g., fzy instead of or in addition to fizzy). Learn to use FlyBase so you always have access to current stock lists (read stocks.doc before trying to search the stock lists). Don't order Bowling Green stocks from Bloomington, and vice versa. Don't order the same stocks from multiple stock centers 'just to be sure'. If you find yourself often ordering one or two stocks a week for several weeks in a row, consider delaying your next order for a week or two so you can order everything you will need for a while at once. For small orders, the processing and packaging time vastly outweighs the stock set up time, and up to 12 stocks can be shipped for the same postage as one. It is very helpful when workers in the same lab pool their orders. We try to identify multiple orders from the same lab and ship them together, but this takes extra time, and we aren't always aware of you lab affiliation. 6. Check the redbook or FlyBase for basic information about a gene or an aberration before calling the stock center for such information. * NO, IT WASN'T YOUR IMAGINATION -- Try to forgive us if you suffered from Kathy's even-crankier-than-usual disposition this fall. We were seriously oversubscribed and biting a head off now and again just felt too good to resist. Our five year renewal application is in, we are once again fully staffed, and spring isn't all that far off, so you should be safe for a while. *** FLYBASE A new release of FlyBase will appear on the server sometime in January. It will be announced on bionet.drosophila. If you don't have local access to the bionet.drosophila discussion group you can subscribe directly by sending an e-mail message to BIOSCI@NET.BIO.NET asking to subscribe to bionet.drosophila. A person will read your message, so it need not be in any specific format. You may post a message to the group by sending your message to DROS@NET.BIO.NET. *** REQUESTS FOR MATERIALS WILD-CAUGHT bb MUTANTS Leonard G. Robbins, Genetics Program, S308 Plant Biology, Michigan State U., E. Lansing, MI 48824-1312. 517-355-0337, FAX/353-1926, 21675MGR@MSU.EDU or 21675MGR@MSU.BITNET. For an attempt to find other instances of Rex, I would appreciate cultures of any wild-caught or spontaneous melanogaster bb mutants. *** MATERIALS AVAILABLE MONOCLONAL ANTIBODY AGAINST EMBRYONIC CHORDOTONAL ORGANS Beate Lichte, Thilo Schneider, and K.-F. Fischbach, Inst. fuer Biologie III, Schaenzlestr. 1, D-79194 Freiburg, Germany. 0761-203-2730, FAX/2745, LICHTE@SUN1.RUF.UNI-FREIBURG.DE or KFF@SUN1.RUF.UNI-FREIBURG.DE. We recently produced (as a "by-product" of our current research) a mouse monoclonal antibody which recognizes exclusively all chordotonal organs of the Drosophila embryo in histochemical staining experiments. The staining is very strong without any background. So, if anyone is interested in using this antibody, e.g. as a marker, please contact us. *** TECHNICAL NOTES PREPARATION OF DNA FROM SINGLE EMBRYOS FOR PCR Maryann Garozzo and Alan C. Christensen, Dept. of Biochemistry and Molecular Biology, Thomas Jefferson U., 233 S. 10th Street, Philadelphia, PA 19107, 215-955-5190, FAX/5393, CHRISTEN@CALVIN.JCI.TJU.EDU. We have adapted the single fly PCR method of Gloor and Engels (DIS 71: 148-149, 1992, and DIN Vol. 1, 1991) to single embryos. We have also slightly modified their procedure for single fly PCR which gives less background in our hands. The ability to use single embryos for PCR allows one to determine the genotype of an embryo following phenotypic analysis or other manipulation. Since there are relatively few good visible phenotypic markers for embryos, polymorphic sequence tagged sites can be used as chromosome markers in individual embryos. Single P element inserts can also be used; in this context they serve as portable sequence tagged sites. The sex of the embryo could also be determined by this method. 1. DNA PREPARATION FROM EMBRYOS. Single embryos are squashed in 10 ul of Gloor and Engels' extraction buffer (10mM Tris pH 8.2, 1mM EDTA, 25mM NaCl, 200ug/ml proteinase K freshly diluted from a frozen 20mg/ml stock). This is most conveniently done in a 0.5 ml microfuge tube, using the pipettor tip to crush the embryo in the buffer. Care should be taken to avoid getting the embryo stuck inside the pipettor tip. The homogenate is incubated at 37[o]C for 30 minutes, then 95[o]C for 2 minutes, then stored at 4[o]C. It is easy to program a thermocycler for these incubations. We typically use 1 ul of this extract in a 15-50 ul PCR using standard conditions, as appropriate to the primers. 2. NOTES ON THE EMBRYOS. We have successfully amplified single copy sequences with this procedure using embryos 12 hours old and older. It also works with first instar larvae. The embryos may be dechorionated or not. If they are dechorionated, we have found (not surprisingly) that the bleach must be thoroughly rinsed off. We have also used this procedure on embryos that have been permeabilized with heptane, immersed in halocarbon oil or stained for programmed cell death with acridine orange (Abrams et al., Development, 117: 29-43, 1993) If the embryos have been in halocarbon oil, we wash the oil off with heptane, although this may not be necessary. None of these procedures appears to interfere with DNA extraction or PCR. We have not attempted the procedure with fixed embryos. 3. MODIFICATIONS OF THE SINGLE FLY PCR PROCEDURE. Generally, the procedure of Gloor and Engels works very well. However, we have occasionally had problems with spurious background bands, and these are often worse when the priming sites are absent in the fly being tested. This problem is alleviated with no loss of the bona fide amplification product by using less fly extract in the PCR. For example, if the fly was homogenized in 50 ul, we use 1 ul in a 50 ul reaction, rather than 1 in 15. Reducing the number of PCR cycles from 30 to 25 also reduces the amount of background with no loss of signal. ***