Thanks from our lab to all those friendly responses to our query.
Herewith again my original request and the promised summary of all
the suggestions  and tips (if you are interested in PDFs or protocolls
or other links - please let me  know, I won't attach it all here cause I
don't want to hog all your mailboxes):

our initial query:
"we are busy with phyolgeographic studies of Scarabs (Coleoptera)
and are struggling to get enough good loci in a microsatellite library
(even using enrichment).
Does anyone have experience with microsatellites in beetles and
would be prepared to share this (especially encountered problems or
special tricks or idiosyncracies)?
Is it possible that Coleoptera (just as Lepidoptera) do not have that
many microsatellite loci?
Could you recommend any other nuclear marker for phylogeographic
and conservation genetic studies in beetles?"

..and now ...the answers:

1) I would say that Coleoptera have plenty of microsatellites, but they
are difficult to find.

I suggest two things.

1- try and get access to the EST libraries of Volger and company, see
attached paper.  Scan these EST seqeunces for microsatellites.
2- move on and try and get nuclear gene markers that are variable
with indels.  I think that this is pretty easy to do.  Again, get the
ribosomal protein genes for Coleoptera, which are now in gene
bank.  Make primers at the 5' and 3' end of these genes and PCR
them
for several individuals.  I imagine you will find some size variation
in the PCR products.  Just use this as your molecular marker.  You
could also try and do Restriction Enzyme digests on these PCR
products that don't have any size variation- to get variation among
individuals in what cuts/does not cut.

See the second attached paper explaining that using nuclear genes
can
be just as informative as microsatellite loci.  Given that you
probably can't get the micorsatellite loci, you certainly can get the
ribosomal proteins to PCR for your species.

best of luck and let me know if you have any questions,

Chris

2) Dear Ute,

my husband was just forwarding me your mail, there are two things:
first it looks as if some beetles really do not have many repetative
sequences in the genome, but that is not easy to estimate. In the lab
I used to work were two students which were isolating mircosats from
beetles (Curculionoidea and Chrysomeloidea), and both had
problems
but at the end they got some. Here in Madrid they use a very fast
PCR
based method that seems to work well for the group of animals they
are working with. If you want I can ask the detailed protocol.
The other thing is that you could try ITS, this is easy to amplify
and for population/subspecies studies not too bad. This might be
worth to try. I have been using this for Lepidoptera and certainly
can find the primer sequence but they should be available in the
literature.

Hope that helps a bit, in case you have some molecular questions
please do not hesitate to ask, I am not very good with the beetles
but the molecular work is fine.

Greetings to Clarke, even though he might not remember me from
the
time in London, since I am not a beetle person.

All the best,
Ali
--
Dr. Alexandra Cieslak
Department Biodiversidad y Biologia Evolutiva
Museo Nacional de Ciencias Naturales
Jose Gutierrez Abascal, 2
28006 Madrid
Spain

3) We have developed primer sets for about 65 gene regions (single-
copy
protein
coding genes) IF you are interested in cDNA work (RT-PCR).  If you
are
uncomfortable working with RNA, we have a few of the same genes
that work
with
genomic DNA.

bernie ball
Duke University

4) Dear Ute,

In response to your message regarding useful nuclear markers in
Coleoptera: I am a phylogeneticist specializing in Auchenorrhyncha
(Hemiptera), however, I worked on a molecular phylogenetic the
Hawaiian
Platynini (Carabidae; Jim Liebherr's group of interest) as a Post-doc.
Attached is a .pdf of the paper resulting from my work with that
group,
in which I sequenced 28S and wingless from the nuclear genome.  I
know
that Jim Liebherr has published a number of other papers on
Platynini
that deal more specifically with the group's phylogeography, as well.

Best of luck!

Jason

Jason R. Cryan, Ph.D.
Director, Laboratory for Conservation and Evolutionary Genetics
New York State Museum
3140 Cultural Education Center
Albany, NY  12230
Phone: (518) 486-2008
E-Mail: jcryan@mail.nysed.gov
FAX: 518-486-2034

5) Hi Ute,
I'm a PhD student working on beetles (Tenebrionidae) in fragmented
systems. I too have found it very difficult to develop good
microsatellite loci as well as other non-microsatelite nuclear markers
for population genetics.
The best I can offer is our paper (attached):
Schmuki C, Blacket MJ, Sunnucks, P (2006) Anonymous scnDNA
markers for
two endemic log-dwelling beetles: Apasis puncticeps and Adelium
calosomoides (Tenebrionidae: Lagriinae: Adeliini). Molecular Ecology
Notes 6, 362-364.
Unfortunately our exhaustive efforts yielded only 5 polymorphic
markers
in total for two species with number of alleles varying between 2 and
10, but the markers successfully detected effects of habitat
fragmentation for the two species.
Good luck with it,
Christina Schmuki.

6) Hello Ute,

we just were working on  P. chalcographus (Scolytinae, Coleoptera)
and had
problems isolating loci. Already 1997 I worked on Ips typographus ,
another scolytid, without success, however not using enrichment. But
since
that time I met several people working on scolytids encountering
similar
problems.

Wolfgang Arthofer did his PhD in my group and he spent some hard
months
and got only some loci which we are trying now to
publish. I/He could send you a pdf of his PhD so that you know what
he
tried. Generally we oberved

- only few loci per library
-repaeted isolation of identical loci
- often short and interrupted motifs.

The ones published for scolytids by other groups are not really
working
well at least for Ips tpyographus - I can send you a paper in press in
Biol J Linn Soc.  Further there was a isolation in chrysomelids but the
authors never published an "bigger" paper -  for me indicating that the
loci are not working that well....

It would be nice if you let us know which answers you receive and
particularly your specific methods and problems - it seems there is a
general problem with isolation from coleopterans - exception
carabids.

If you have questions don't hesitate to contact me or Wolfgang.

Best wishes
Christian

Christian Stauffer, Department of Forest & Soil Sciences, Institute of
Forest
Entomology, Forest Pathology & Forest Protection; BOKU, University
of
Natural Resources & Applied Life Sciences, 1190 Vienna, Austria

Tel: +43 1 3686352/25 Fax: /97 http://ifff.boku.ac.at
email: christian.stauffer@boku.ac.at

7) Dear Ute,

I have experience with microsatellite screening in Scolytidae, and
also
experienced troubles... We had to use an enrichment protocol, and
even
doing so the proportion of positive clones was very low. We even had
to
forget about microsatellites in one species (the few we found were
monomorphic or impossible to interpret). I think the best strategy is to
enrich once or twice (but then be aware that you'll find redundant
clones in the positive clones you'll find) and clone as many colonies
as
possible because you'll find a low number of loci actually containing a
microsat.

The situation cannot be generalized to Coleoptera though. Carabid
species seem to be OK (genera Carabus and Chrysocarabus were
ok), as
well as some Curculionids (We had no problem with the genus
Pissodes). I
am sorry to hear that Scarabeids have also low microsat contents.

You could try AFLP, but you cannot answer all questions using
those...

Good luck,

Carole

--

Carole Kerdelhu�
INRA, UMR BioGeCo
69 Route d�Arcachon
F-33612 Cestas Cedex, France.
Tel : +33 5 57 12 28 58
Fax : +33 5 57 12 28 81
http://www.pierroton.inra.fr/biogeco/entomologie/personnel/Kerdelhue
/Kerde
lhueperso.html

8) Dear Ute,

I am afraid I do not have any answers to your questions.  However,
as I
work on species-level evolution and molecular genetics in beetles
(mostly
chrysomelid leaf beetles), I would be interested in what information
you
receive from your request.  It would be great if you could post these
responses on the evoldir as other authors have done.

Although I have not yet worked with microsatellites, we have recently
been
having considerable success using AFLPs to study genome-level
genetic
differentiation.  The technique has proved straightforward and the
data
are highly informative.

Good luck with your work.

Sincerely,

Dan

Daniel  J. Funk
Assistant Professor
Dept. of Biological Sciences
Vanderbilt University

9) Dear Ute,

My lab has had experience getting microsatellite loci from water
beetles
(Dytiscidae) without too many problems that I am aware of. Michelle
Guzik
(Michelle.Guzik@adelaide.edu.au), a post doc with me, has been
doing the
lab work if you would like to get more details of her protocol (a
microsatellite enrichment technique) and the types of loci she has
isolated. I think there are a number of other beetle microsatellite loci
cloned (see Molecular Ecology Notes).

regards,

Steve

Dr Steve Cooper
Evolutionary Biology Unit
South Australian Museum

10) Dear Ute.
	I found short time ago this paper talking about the
singularity of the microsats in Lepidoptera.
	I attach it, and hope can be interesting for you and your
project.

Best regards.
Joaquin Munoz.
--

Un buen lugar para descansar y disfrutar de la naturaleza y
gastronomia:
http://www.laposadademontellano.com/

Joaquin Munoz Garcia
(PhD. Student)
Laboratory of Molecular Ecology.
Estacion Biologica Donana (CSIC).
Pabellon del Peru.
Avda. Maria Luisa, s/n.
Apartado de Correos 1056.
41013-Sevilla.
Spain.
Phone: #34-95-4232340 (Office ext. 236; Lab. ext. 221)
Fax: #34-95-4621125
http://www.ebd.csic.es
http://www2.ebd.csic.es/lem/Personal/Quini.htm

11) Hi Ute,

         In reference to your post on Evoldir, we developed a panel of
microsatellites in southern pine beetle (Dendroctonus frontalis) using
a
common enrichment protocol and had no difficulties.  I know
colleagues who
have developed microsats in similar species with varying levels of
success.

Regards,

         Ed Heist

Edward J. Heist, Ph.D.
Southern Illinois University at Carbondale
Fisheries and Illinois Aquaculture Center
Life Sciences II, 1125 Lincoln Drive
Carbondale, IL 62901-6511
Voice: (618) 453-4131
Fax: (618) 453-6095
email: edheist@siu.edu
web: http://www.science.siu.edu/zoology/heist/index.html

12) Hello Ute,
My name is Elena and I am working at the National History Museum
of
Madrid. I am actually writing you in behalf of  Alexandra Cieslak,
(see email below).  Here I send you some protocols I've been using
for the isolation of microsats. I hope they will help you. Please,
feel free to ask me if you have any other question. Good luck!

Cheers,
Elena

13) Hi Ute,

Well, i cant help with the microsats, but on the question of other
nuclear
markers, did you see the paper by Pons et al. in sys. biol. about the
MP20
introns? They used Cicindela (adephagan Coleoptera), but the
primers
were
designed from a broader sample of beetles.

I was just playing with generating a matrix of insect MP20, to look at
the
broader evolution of the gene. There is plenty of data from this gene
if
you
want to redesign primers that might be more specific to scarabs. The
closest
taxa to scarabs with data for this gene is Georissus (Staphylinid), but
there
is lots of comparative data from elateriform or cucujiform beetles.

The paper on MP20 as a marker in adephaga is available here.
http://72.14.203.104/search?q=cache:KYFvWyeZlM4J:hydrodictyon.e
eb.uconn.ed
u/courses/systematicsseminar/restricted/Pons_et_al_2004_SystBiol_
53_554-57
0.pdf+MP20+cicindela&hl=en&gl=us&ct=clnk&cd=5

Let me know if you want to talk about available MP20 data and
modifying
the
primers etc.

Best of luck
stuart

Dr. S. Longhorn, MSc PhD
PostDoctoral Researcher
Mitogenomic Analysis of the Arachnid Orders
Rm. 105 Graduate and faculty office,
SB 1; Dept. of Biology
Portland State university
Portland, OREGON, 97207
Lab: TEl:001-503-725-3855
Email: sjl197@hotmail.com or stul@pdx.edu

WORK CURRENT: http://web.pdx.edu/~smasta/
WORK PREVIOUS:
http://www.bio.ic.ac.uk/research/apvogler/vogler.htm

14) Dear Ute,

Someone has just forwarded me your enquiry about markers for
beetles, I
must somehow have missed your message.  I developed
microsatellite
markers for ladybird beetles (primer note attached), so have some
experience of this.  Are you still in need of help or have you sorted
your problems out?  I'm not sure if there's anything I can do, but if
there is then please do let me know.

Best wishes

Penny Haddrill

so - once again, thank you all you wonderful co-scientists,
and may the (nuclearDNA) force be with you!!!
Ute

Ute Kryger (PhD)
Scarab Research Group
Department of Zoology & Entomology
University of Pretoria
Pretoria 0002 - RSA
Fax +27 12 362 5242
Phone +27 12 420 5343

Ute Kryger <ukryger@zoology.up.ac.za>