7:30 Coffee in Exhibit Hall

Creating a Higher Throughput Laboratory

8:00 Chair’s Remarks
Dr. Phillip Danielson, Associate Professor, Biological Sciences, University of Denver

8:05 The Development of Highly Integrated and Automated Forensic Specimen Processing Platforms
Dr. Thomas Scholl, Director, Product Development, Myriad Genetics, Inc.

8:35 Expert Systems for Forensic Sample Analysis
Dr. Stephen Sherry, Staff Scientist, NCBI/NIH

9:05 New Methods Created for High Throughput Casework Analysis
Dr. James W. Schumm, Chief Scientific Officer, Vice President, The Bode Technology Group
To ease the national backlog of crime scene evidence requiring DNA analysis, we have adopted methods that support high throughput processing of casework samples. In particular, we have developed a new device for simplified reference sample collection, new protocols for a rapid pre-screen for Y chromosome DNA and microplate-compatible human-specific quantification, and a computer program to convert Genotyper output to court-ready documents. Recently, we have been working in conjunction with the FBI Laboratory to implement robotic automation of key components of sample handling for STR-related procedures.

9:35 Automated Filtration-Based DNA Preparation for Sexual Assault Cases
Dr. Alex M. Garvin, Director of Molecular Diagnostics, Bureco, AG
The method used for isolating sperm DNA from sexual assault cases has not changed significantly in the 18 years since the selective lysis protocol was first published. This method involves centrifugation and careful removal of supernatant and as result is difficult to automate. A simple alternative for collecting and washing the intact sperm present after selective lysis is to use a filter. I have identified a special type of filter and have designed a protocol for its use that works well for this application. Since only pipetting and vacuum drived filtration steps are required, the process can easily be automated in a 96 well format using existing robotic work stations.

10:05 Automated PCR Setup for Casework Samples as Part of a Total Automated System
Mrs. Gabriela Saldanha, Product Manager, Genetic Identity, Promega Corporation
This presentation will discuss a flexible automated PCR setup system that works over a 50-fold range of initial DNA concentration and is ideally suited for casework samples. The system uses DNA concentration information and user-defined volumes to calculate and dilute DNA to a set concentration. PCR master mix and the normalized DNA are then placed in PCR tubes or plates. In addition, improvements for automated DNA purification and human-specific quantitation using the same robotic instrument as PCR setup, the Beckman Coulter Biomek® 2000 Workstation, will be described. This modular approach provides quicker access to automation, minimizes cost, and provides the flexibility to update and substitute modules as desired.

10:35 Refreshment Break, Poster and Exhibit Viewing
(Last Chance to View Posters and Exhibits)

11:15 Comparison of mtDNA Hair Extraction Techniques
Mr. Mark Kavlick, Biologist, CTFSRU, FBI Laboratory
Traditional methods to extract and purify mtDNA from hair shafts typically include a grinding step followed by organic extraction and membrane filtration. Several commercially available kits are now available for hair mtDNA extraction, which eliminate these steps and offer convenience, speed, and fewer manipulations. However, it is not known whether the quality of mtDNA preparations obtained by these new methods are comparable to the traditional method. These kits were evaluated against the standard method using a variety of donor hair shafts and lengths for their ability to yield sufficient quantities of mtDNA for successful amplification as well as producing high quality sequence data.

11:45 Human mtDNA Mixture Separation by Denaturing High-Performance Liquid Chromatography
Dr. Phillip Danielson
Mitochondrial DNA (mtDNA) sequencing can provide crucial information to forensic investigators when the quantity and quality of DNA would otherwise be limiting. Because mtDNA amplicons used for forensic analyses are identical or nearly identical in length, however, it is virtually impossible to determine the mitotypes of individual contributors to a DNA mixture by direct sequencing. The methods currently available to separate the individual contributors to a mixture (e.g., subcloning) are not only time consuming but are also prone to the generation of artificial sequence variants due to the less-than-perfect fidelity of Taq DNA polymerase. Denaturing High-Performance Liquid Chromatography (DHPLC) employs temperature modulated heteroduplex formation to achieve sequence-specific separation of the individual components of a mixture. DHPLC can be used as an accurate method for the rapid, low-cost separation of both natural (heteroplasmic) and situational (multi-contributor) mixtures without the need for reamplification or excessive manipulation prior to DNA sequencing.

12:15 DNA Witness
Mr. Zach Gaskin, Technical Director of Forensic Genomics, DNAPrint Genomics, Inc.
Five murdered and sexually assaulted women from around the Baton Rouge area were found to have a common foreign DNA donor through STR DNA analysis. The genetic profile of the donor from the crime scene specimens could not be found in a database. With no suspects, the Louisiana Task Force set out to dragnet suspects from the local community. Eyewitness accounts of a Caucasian male acting suspicious near the crime scenes focused the efforts of the task force in what would later prove to be the wrong direction. In February 2003, the Louisiana State Police Crime Lab contracted the services for DNA WITNESS testing after the dragnet DNA testing produced no “hits” from the more than 700 individuals tested. Our results indicated the killer to be 85% Sub-Saharan African and 15% Native American, and two months after receiving this information, the task force had an African American male in custody that matched the STR profile found at each crime scene. This SNP-based DNA test for the determination of an individual’s Biogeographical Ancestry (BGA) has been utilized for genealogy enthusiasts, adopted individuals, and persons wanting to prove Native American affiliation. This presentation will provide information on the scientific foundation of the test and how it can and should be applied in modern forensics.

12:45 Close of Conference