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Modified Method for Purification of Biological Samples Collected on FTA Cards for STR Analysis

NCJ Number
213439
Journal
Journal of Forensic Identification Volume: 56 Issue: 2 Dated: April 2006 Pages: 222-231
Author(s)
Mark Barash; Moshe Shpitzen; Ron Gafny; Ashira Zamir
Date Published
April 2006
Length
10 pages
Annotation
This study compared the recommended protocol of DNA purification and amplification from FTA cards to a simplified method the authors developed in the laboratory.
Abstract
The method developed by the authors reduced the time required to perform the washing and amplification steps, reduced the amount of PCR reagents required, and used distilled water rather than the FTA reagent. This method would, therefore, be useful in high-volume laboratories. The proposed protocol could also be automated and used for processing a large number of samples for DNA databases. The modified protocol consists of a single wash in double-distilled water for 7 minutes and performing the PCR reaction in the same tube. The authors found that a PCR that uses 3.5 ml of primers, 0.4 ml of Taq Gold polymerase, combined with the reduced size of the sample disk (1 mm) and alternative washing and amplification protocols are the best conditions for obtaining a complete DNA profile from the FTA card carrying blood samples, using the AmpFLSTR SGM Plus PCR kit. This article describes in detail the isolation of DNA from FTA cards by purifying DNA according to the manufacturer's protocol. It then describes the alternative washing protocol developed by the authors. Also described are the DNA quantitation, amplification, and analysis. 1 figure and 7 references