NCJ Number
196327
Journal
Journal of Forensic Sciences Volume: 47 Issue: 4 Dated: July 2002 Pages: 811-818
Date Published
July 2002
Length
8 pages
Annotation
This paper describes an improved method for post-PCR purification for mtDNA sequence analysis.
Abstract
Mitochondrial DNA (mtDNA) analysis of forensic samples typically is performed when the quantity and quality of DNA are insufficient for nuclear DNA analysis or when maternal relatives may be the only reference source. The current procedure requires purification of the amplicons by centrifugal filtration after PCR and prior to cycle sequencing. Because this method involves several manipulations, alternate cleanup procedures were investigated. When the yield of purified amplicons, quality of the sequence profile, and ease of assay were evaluated, ExoSAP-IT reagent for post-amplification purification was chosen to replace the filtration method. It provides consistent, efficient recovery of amplified DNA; the amount of DNA recovered exceeds that obtained by use of a Microcon-100 filtration device; and the quality of subsequent sequencing profiles is comparable for all purification methods evaluated whether the amplified DNA is analyzed immediately or after storage of up to 2 months. ExoSAP-IT treatment is amenable for forensic-type samples, as demonstrated on evidentiary-type samples consisting of whole blood, hair, and a tooth. Figures, tables, references