PROJECT ANT:
BRINGING MICROSATELLITE TECHNOLOGY
TO THE CLASSROOM
We have successfully used microsatellite primers developed by our mentor, Daniella Calcagnotto, to analyze samples of Piaractus mesopotamicus which she collected in Brazil. We envision using this technology and related analytical techniques to do ecological and population genetics studies with our students. We believe that teaching these techniques in order to do science will be much more powerful than simply teaching on a demonstration basis. This project incorporates the science-as-inquiry vision of the National Science Education Standards (1995) and focuses on learning subject matter disciplines within the context of inquiry and technology. Here we outline a project we believe will work in the classroom and which we hope to develop and test over the next year.
Choosing an Organism
We have chosen to study ants. "The neglect of ants in science and natural history is a shortcoming that should be remedied, for they represent the culmination of insect evolution" (Holldobler and Wilson, The Ants) and the study of their social organization is endlessly fascinating. They are readily found and collected in all environments, populations can be established and maintained in the classroom and their study creates an opportunity for students to interact with their immediate environment.
It is critical for classroom study that primers for the organism in question, or a closely related organism, be available. Primers identify microsatellite loci for amplification and identification. Generation of primers requires identification of microsatellite sites, a laborious process involving screening of a genomic library, see Calcagnotto et als, Isolation and characterization of microsatellite loci in Piaractus mesopotamicus and their applicability in other Serrasalminae fish, Molecular Ecology Notes (2001) 1, 245-247, a procedure beyond the scope of a classroom study.
Sequences of microsatellite primers may be found by going to the web site maintained by the National Center for Biotechnology Information (NCBI) and doing a nucleotide search for "microsatellite" with the name of the organism or group in question. We located microsatellite primers for two well-studied ant species which are common in the United States, Lepothorax acervorum (northern regions) and Linepithema humile (southern regions). Once the primer sequence is known, the primer can be made-to-order commercially. If a primer is not available for a particular species, primer locations for closely related groups may work, Rico et als, 470 million years of conservation of microsatellite loci among fish species, Proc. R. Soc. Lond. B (1996) 263, 549-557, and may be tested by a PCR reaction on an available sample.
Initiating the Project
This year we plan to survey the vicinity of our schools for colonies of L. acervorum, L. humile and closely related species with the objective of conducting prototype microsatellite experiments to study population genetics and other questions. Our students will classify the ants, GPS-map the colonies, study the natural history of the ants and initiate collection for prototype experiments during the summer. These activities will present a host of educational activities for our students. Depending on progress and availability of equipment, we may do DNA extractions and tests of primer efficacy during the school year.
Prototype Microsatellite Analysis Experiments and Documentation
Our demonstration project this summer with pre-existing fish samples and primers has convinced us of the feasibility of doing this type of project within the classroom context. We need to do additional work to ensure that the project is classroom-ready (i.e., the project must be safe, be organized so that discrete segments can be completed within the school year, have complete materials and equipment lists, have protocols for use by students and teachers, have appropriate learning resources and assessment tools and have a high probability of producing meaningful science). We hope to spend next summer doing a complete demonstration set of experiments with the sample organisms collected by our students and creating protocols for use by other teachers.
Partnerships
There are two critical and expensive pieces of equipment required for this project, a thermal cycler for the PCR reactions and a sequencer to do the gene scans necessary for microsatellite analysis. Schools will have to establish partnerships with universities or local biotechnology firms in order to have access to sequencing facilities. This is an outstanding opportunity to forge important relationships between schools and their communities. It would be desirable but not essential for schools to have thermal cyclers on site. BioGENEator, a programmable thermal cycler kit, is available from Cold Spring Harbor labs for $800, and other moderately-priced options are available.
Further Information
For further information, contact David Hinden, Harvard-Westlake School, N. Hollywood, CA
or Phillip Moore, Fitchburg HS, Fitchburg, MA