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January 01, 2009 (Vol. 29, No. 1)

High-Throughput Automation of DNA Extraction

Generating Amplifiable DNA for Nucleic Acid Analysis

  • High-Throughput Applications

    Click Image To Enlarge +
    Figure 2. Set up of automated DNA extraction on the epMotion 5075 Liquid Handler for forensic FTA punches in 96-well format.

    High-throughput EA1 TCE has many applications—from basic research, drug discovery and development, clinical research and diagnostics, to agriculture, animal and livestock, and forensics. Experiments employing forensicGEM illustrate the method’s high-throughput automation capabilities.

    Forensic laboratories worldwide conduct DNA profiling of individuals for criminal DNA database development. These labs process thousands of samples, comparing database profiles against crime-scene DNA to generate cold hits. Often having a significant backlog of samples to process, many forensic laboratories seek streamlined automation in DNA extraction and STR analysis. The use of spin columns or magnetic beads is generally expensive and difficult to automate for high-throughput application.

    In one set of experiments, an Eppendorf epMotion® 5075 robotic system, fitted with twin Peltier blocks and a thermal cycler (Figure 2) was used for forensicGEM DNA extraction of FTA card punches in 96-well format, followed by STR analysis using industry-standard STR kits. Data demonstrated (Figure 3) that this high-throughput automation of DNA extraction and STR analysis was ideally suited for forensic application and can streamline sample processing and analysis. The extraction process takes approximately 35 minutes with 4–5 minutes of technician time required initially for reaction preparation.

  • Conclusion

    Click Image To Enlarge +
    Figure 3. (A) Multiplex STR-PCR (STR Identifiler® PCR Kit, Applied Biosystems) analysis obtained from DNA extracted from liquid blood samples using forensicGEM.

    Traditional DNA sample preparation for genetic analysis assays can be a time-consuming, costly, and labor-intensive process. ZyGEM’s DNA-extraction technology using EA1 involves a temperature change without need for wash or elution steps. This simplicity lends itself to automated, high-throughput application using most existing liquid-handling systems and can simplify laboratory workflow from DNA extraction to nucleic acid analysis.

  • Click Image To Enlarge +
    Figure 3. (B) TaqMan® Real-time quantitative PCR trace

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