Drosophila Information Newsletter Vol. 13

Kathy Matthews matthewk at fly.bio.indiana.edu
Fri Dec 31 19:01:01 EST 1993


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 at 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 at INDIANA.EDU
CTHUMMEL at HMBGMAIL.MED.UTAH.EDU          MATTHEWK at INDIANA.BITNET
***

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***
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 at 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 at 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 at 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 at 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 at 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 at MSU.EDU or 21675MGR at 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 at SUN1.RUF.UNI-FREIBURG.DE or
KFF at 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 at 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.
***



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