Volume 41, Issue 4 (July 1996)

    Chemiluminescent Detection of RFLP Patterns in Forensic DNA Analysis

    (Received 22 May 1995; accepted 7 November 1995)

    CODEN: JFSOAD

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    Abstract

    DNA testing by restriction fragment length polymorphism (RFLP) analysis is an extremely important technique used in forensic science laboratories. While RFLP testing is a highly informative method, it traditionally has had several disadvantages. It is time consuming and involves work with radioactive phosphorous. A detection method that is faster and safer than isotopic detection is presented. Various membranes, fixation methods and transfer procedures were evaluated for DNA retention and sensitivity using alkaline phosphatase conjugated oligonucleotide probes and a chemiluminescent substrate. Blood samples and evidentiary material from forensic casework were analyzed by both chemiluminescent and isotopic detection. Results of each method were compared for pattern appearance, band size, and composite profile frequency. The chemiluminescent system had very good sensitivity, detecting 3–25 ng K562 DNA. Most patterns developed by both methods appeared the same. The variation observed between band sizes and frequency estimates generated by each method was as expected for an inter-gel comparison. The chemiluminescent detection procedure described here is suitable for use in forensic casework.


    Author Information:

    Johnson, ED
    Forensic chemists, U.S. Army Criminal Investigation Laboratory, Fort Gillem, GA

    Kotowski, TM
    Forensic chemists, U.S. Army Criminal Investigation Laboratory, Fort Gillem, GA


    Stock #: JFS13957J

    ISSN: 0022-1198

    DOI: 10.1520/JFS13957J

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    Author
    Title Chemiluminescent Detection of RFLP Patterns in Forensic DNA Analysis
    Symposium , 0000-00-00
    Committee E30