David Micklos, John Kruper, Scott Bronson, and Matthew Christensen 
DNA Learning Center, Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, New York 11724 
micklos@cshl.org 

The DOE Program introduces high school biology faculty to a laboratory-based unit on human DNA polymorphisms, which provides a uniquely personal perspective on the science and ELSI aspects of the Human Genome Project. Thus far, 170 high school faculty have participated in eight three-day workshops held in Atlanta, Boston, Denver, Los Angeles, New York, Richmond, Salt Lake City, and San Francisco. Participants learn simplified lab techniques for amplifying three types of DNA polymorphisms: an Alu insertion, a VNTR repeat, and point mutations (SNPs) in the mitochondrial (mt) control region. These polymorphisms illustrate the use of DNA variations in disease diagnosis, forensic biology, and identity testing - and provide a starting point for discussion of the uses and potential abuses of genetic technology. 

During the past year, we developed ready-to-use teaching kits to support the three human PCR experiments introduced in the DOE workshop. The kits, which are now available through Carolina Biological Supply Company, incorporate a three-part PCR chemistry that greatly simplifies reaction set-up and improves reproducibility. Template DNA (obtained from cheek or hair cells) is added to primer mix and a freeze-dried reagent pellet (containing Taq polymerase, deoxynucleotide triphosphates, and buffer). Loading dye is incorporated in the primer mix, allowing amplified reactions to be immediately loaded for gel electrophoresis. 

We also provided proof of concept for a Sequencing Service to process mt DNA samples submitted by teachers around the country. During each DOE workshop, participants amplified the mt control region, and the samples were returned to the CSHL Genome Sequencing Center for cycle sequencing. The completed sequences were then posted at the DNALC's WWW site (http://www.dnalc.org/) in a Student Sequence Database, which currently contains 350 sequences. This process was replicated with 59 student samples submitted, by mail, from teachers in New York, Maryland, Utah, and Virginia. A dedicated DNA sequencer to support student sequencing is included in the capital budget for a 6,000 square foot BioMedia addition to the DNALC, on which construction will begin next year. 

We have continued to develop step-by-step Internet templates that allow students to use their own polymorphism data to solve cases illustrating key principles of genomic biology. In one case, the Student Allele Database facility is used to compare students' Alu insertion data with data from world populations. In another case, multiple sequence alignment is used to compare a student and reference mt DNA sequences to determine whether Neanderthal hominids were our direct ancestors. We are currently developing Bioforms that further simplify data submission/presentation - allowing students to focus on the biological question at hand, rather than being overwhelmed navigating complex forms at Internet genome servers. The initial Bioform allows students to analyze mt DNA sequences to identify the remains of the Romanov family and determine if Anna Anderson was the missing princess Anastasia.