automated instrument for human STR identification: Design, characterization, and experimental validation

• Published in: Electrophoresis 2010-10, Vol.31 (21), p.3510-3517
• Main Authors: Hurth, Cedric, Smith, Stanley D, Nordquist, Alan R, Lenigk, Ralf, Duane, Brett, Nguyen, David, Surve, Amol, Hopwood, Andrew J, Estes, Matthew D, Yang, Jianing, Cai, Zhi, Chen, Xiaojia, Lee‐Edghill, John G, Moran, Nina, Elliott, Keith, Tully, Gillian, Zenhausern, Frederic
• Format: Article
• Language: English
• Subjects: Automated; Cheek; Cost analysis; Deoxyribonucleic acid; DNA - chemistry; DNA - isolation & purification; DNA STR typing; Electrophoresis; Electrophoresis, Capillary - methods; Forensic engineering; Forensic Genetics - methods; Human; Humans; Instrumentation; Microfluidic Analytical Techniques - instrumentation; Microfluidic Analytical Techniques - methods; Microfluidics; Microsatellite Repeats; Mouth Mucosa - cytology; Polymerase Chain Reaction - methods; Reproducibility of Results; Sequence Analysis, DNA - instrumentation; Sequence Analysis, DNA - methods; Temperature
• ISSN: 0173-0835; 1522-2683; 1522-2683
• DOI: 10.1002/elps.201000305

The microfluidic integration of an entire DNA analysis workflow on a fully integrated miniaturized instrument is reported using lab‐on‐a‐chip automation to perform DNA fingerprinting compatible with CODIS standard relevant to the forensic community. The instrument aims to improve the cost, duration, and ease of use to perform a “sample‐to‐profile” analysis with no need for human intervention. The present publication describes the operation of the three major components of the system: the electronic control components, the microfluidic cartridge and CE microchip, and the optical excitation/detection module. Experimental details are given to characterize the level of performance, stability, reliability, accuracy, and sensitivity of the prototype system. A typical temperature profile from a PCR amplification process and an electropherogram of a commercial size standard (GeneScan 500™, Applied Biosystems) separation are shown to assess the relevance of the instrument to forensic applications. Finally, we present a profile from an automated integrated run where lysed cells from a buccal swab were introduced in the system and no further human intervention was required to complete the analysis.