LabAutomation 2006 - SLAS

Where Laboratory Technologies Emerge and Merge
3:30 pm Monday, January 23, 2006 Track 5: Frontiers Beyond BioPharma Room: Sierra/Ventura
Wyndham Palm Springs Hotel
Amy Herr
Co-Author
Sandia National Labs
Victor C. Rucker
Livermore, California
aeherr@sandia.gov
Sandia National Labs
Rapid DNA Fragment Sizing Using Ultra Sensitive Flow Cytometry
DNA fragment sizing is arguably one of the most common measurements performed in today’s biochemical//biomedical laboratories.
Although significant advancements in capillary-based sizing method have been made over the past decade, gel-based electrophoresis
techniques still dominate the DNA sizing arena, especially for large fragments. We have developed an entirely new approach for DNA
fragment sizing based on ultra sensitive flow cytometry. We target fragments in the size range of 1000 base pairs (bp) to 1 Mbp and
have the advantage of requiring extremely small quantities of analyte. We have targeted this technique to sizing of DNA fragments from
restriction fragment length polymorphism (RFLP) digests of bacterial genomes for species identification and strain typing. We present direct
comparisons of our flow-based technique with the “gold standard” pulse field gel electrophoresis (PFGE). Overall our technique is faster
and requires less material than PFGE. Applications of this technique include homeland security, public health, and law enforcement.
4:00 pm Monday, January 23, 2006 Track 5: Frontiers Beyond BioPharma Room: Sierra/Ventura
Wyndham Palm Springs Hotel
Cynthia Bruckner-Lea
Co-Author(s)
Pacific Northwest National Laboratory
R.M. Ozanich
Richland, Washington
B.P. Dockendorff
cindy.bruckner-lea@pnl.gov
M.G. Warner
T.M. Straub
J.W. Grate
Automated Sample Preparation and Detection of Pathogens in Environmental Samples
Methods for the automated purification and concentration of cells, nucleic acids, and proteins are critical to enable the trace detection
of biological analytes in environmental samples. The BEADS platform (Biodetection Enabling Analyte Delivery System) utilizes derivatized
microbeads to capture the analytes of interest, while washing away the sample matrix materials that can interfere with detection. This
presentation will highlight two BEADS system configurations. The first system is configured for the automated sample preparation of
nucleic acids for pathogen identification. This system includes automated cell capture from large sample volumes (milliliters and larger),
followed by flow-through polymerase chain reaction for DNA amplification, and microarray detection. This configuration enables the
detection of only 10 cells/mL in environmental samples. The second system configuration includes on-column fluorescence detection of a
sandwich immunoassay designed for toxin detection. The automated on-column immunoassay enables the rapid detection of toxins in less
than 5 minutes.
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