Jenifer Lindsey, PhD
Unit Chief, DNA Analysis Unit, FBI Laboratory, Washington, DC
ABSTRACT
The use of RFLP analysis is widely accepted in the forensic community as a powerful tool for the characterization of body-fluid stains. In December of 1988, the DNA analysis Unit (DNAU) of the FBI Laboratory implemented the RFLP analysis of casework specimens. Generally accepted to have a high discrimination potential, the success of this technique is dependent on the extraction of sufficient quantities of high quality DNA, thus limiting the types of suitable body fluid stains. Use of 32P-labeled VNTR probes also lengthens turn-around time of results. The initial RFLP protocol used in the DNAU has remained largely unchanged since 1990. This presentation will address validation and implementation of recent changes to this protocol including detection by chemiluminescence, the use of BRL molecular sizing standards, the reduction in EtBr and the alteration of electrophoretic conditions.
The addition of various PCR-based techniques has allowed characterization of forensic serological stains previously deemed unsuitable for RFLP analysis. Current systems used by the DNAU are the Amplitype HLA-DQa and Amplitype™ PM (polymarker) systems. This presentation will address the implementation of analysis of the D1S80 locus multiplexed with the amelogenin locus. Alternative methods of separation and detection will also be discussed.
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