ISSN: 0022-1198
CODEN: JFSCA
Published Online: 1 July 2002
Page Count: 8

An Improved Method for Post-PCR Purification for mtDNA Sequence Analysis
Budowle, B
Federal Bureau of Investigation, VA

Fisher, CL
Federal Bureau of Investigation, VA

Hares, DR
Federal Bureau of Investigation, VA

Lawrence, HS
Federal Bureau of Investigation, VA

Dugan, KA
Federal Bureau of Investigation, VA

(Received 16 February 2002; accepted 6 February 2002)

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.

Keywords:
forensic science, mitochondrial DNA, polymerase chain reaction, DNA sequencing, ExoSAP-IT(tm)

Paper ID: JFS15459J
DOI: 10.1520/JFS15459J
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