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Direct STR Amplification From Whole Blood and Blood- or Saliva-Spotted FTA Without DNA Purification

NCJ Number
222344
Journal
Journal of Forensic Sciences Volume: 53 Issue: 2 Dated: March 2008 Pages: 325-341
Author(s)
Su Jeong Park Ph.D.; Jong Yeol Kim M.S.; Young Geun Yang Ph.D.; Seung Hwan Lee Ph.D.
Date Published
March 2008
Length
7 pages
Annotation
This report describes a new method for direct short tandem repeat (STR) amplification that uses a newly developed direct polymerase chain reaction (PCR) buffer, AnyDirect, which can amplify STR loci from whole blood and blood-spotted or saliva-spotted FTA cards without DNA purification.
Abstract
Regarding concordance with purified DNA, all DNA profiles from blood and blood spots from 50 individuals matched fully; however, regarding saliva spots, poor amplifications were sometimes observed, suggesting that this issue should be studied further. This system had broad compatibility with various DNA polymerase and STR kits. The report also concludes that direct PCR can be applied to old spots, which indicates that the system should be useful for fast DNA typing when there is no need to extract and store DNA, as well as when high-throughput is required. In order to be used actively in forensic case work, further validation studies are required for various kinds of forensic stains and materials that might contain strong PCR inhibitory factors. The autosomal and Y chromosomal STR loci were analyzed for whole blood or saliva spots from 50 random volunteers followed by comparison of the results with those of corresponding purified DNA. In addition to a description of blood and saliva specimens, the description of materials and methods addresses STR amplifications and fragment analysis. The AnyDirect PCR buffer is composed mainly of zwitterionic buffer and/or nonreducing carbohydrates, so as to overcome the PCR inhibitory effects and dNTP's. 3 tables, 5 figures, and 10 references