Volume 47, Issue 4 (July 2002)

    An Improved Method for Post-PCR Purification for mtDNA Sequence Analysis

    (Received 16 February 2002; accepted 6 February 2002)

    Published Online: July

    CODEN: JFSOAD

      Format Pages Price  
    PDF 8 $25   ADD TO CART


    Abstract

    Mitochondrial DNA (mtDNA) analysis of forensic samples typically is performed when the quantity and quality of DNA are insuf-ficient for nuclear DNA analysis or when maternal relatives may be the only reference source. Many of the steps required in the analytical process are both lengthy and labor intensive. Therefore, improvements in the process that reduce labor without compromising the quality of the data are de-sirable. The current procedure requires purification of the amplicons by centrifugal filtration after PCR and prior to cycle sequencing. Because this method requires several manipulations to perform, alternate cleanup procedures were investigated. These include the use of 1) Qiagen QIAquick PCR Purification columns, 2) Concert Rapid PCR Purification columns, and 3) ExoSAP-IT(tm) reagent. When the yield of purified amplicons, qual-ity of the sequence profile, and ease of assay were evaluated, the use of ExoSAP-IT(tm) reagent for post-amplification purification was chosen to re-place the filtration method.


    Author Information:

    Budowle, B
    Federal Bureau of Investigation, Quantico, VA

    Fisher, CL
    Federal Bureau of Investigation, Quantico, VA

    Hares, DR
    Federal Bureau of Investigation, Quantico, VA

    Lawrence, HS
    Federal Bureau of Investigation, Quantico, VA

    Dugan, KA
    Federal Bureau of Investigation, Quantico, VA


    Stock #: JFS15459J

    ISSN: 0022-1198

    DOI: 10.1520/JFS15459J

    ASTM International
    is a member of CrossRef.

    Author
    Title An Improved Method for Post-PCR Purification for mtDNA Sequence Analysis
    Symposium , 0000-00-00
    Committee E30