Comparison of methods for the extraction of DNA from formalin-fixed paraffin-embedded tissues for human identification

Abstract

Formalin-ˆxed para‹n-embedded (FFPE) tissues are one of the most eŠective tools for human identiˆcation in forensic science. To evaluate the eŠectiveness for human identiˆcation, we compared the following six methods for the extraction of DNA from FFPE tissues; EZ1 DNA Investigator Kit, ReliaPrep FFPE gDNA Miniprep System, DNA Isolator PS-Rapid Reagent, QIAamp DNA FFPE Tissue Kit, NucleoSpin DNA FFPE XS and phenol-chloroform method. DNA extracted using each method from two FFPE tissue blocks were quantiˆed by three diŠerent DNA quantiˆcation methods (Quant-it dsDNA HS Assay Kit and quantitative PCR based on 207 bp and 98 bp regions in the D17Z1 locus), and short tandem repeat (STR) typing using AmpFlSTR Identiˆler Plus PCR Ampliˆcation Kit was conducted.All extracts from the FFPE tissues were highly degraded, and large diŠerences were observed among total DNA yields determined by these three quantiˆcation methods. DNA obtained using QIAamp DNA FFPE Tissue Kit had the highest total DNA yield among all the quantiˆcation methods. In STR typing with 1 ng DNA template estimated by quantitative PCR based on 98 bp region in the D17Z1 locus, the greatest number of detected alleles was observed using QIAamp DNA FFPE Tissue Kit. In contrast, EZ1 DNA Investigator Kit and DNA Isolator PS-Rapid Reagent were inferior to the other methods in terms of the DNA yield and the number of detected alleles via STR typing.In conclusion, QIAamp DNA FFPE Tissue Kit was the most eŠective method

Most Common Medico-Legal Autopsy-Related Human and Nonhuman Biological Samples for DNA Analysis

“…DNA isolation from FFPE tissue is a decisive step toward a successful examination. Recent advances in technological developments have made generation of genetic information of DNA and RNA from these kinds of challenging samples possible [110][111][112][113][114].…”

Forensic DNA Technological Advancements as an Emerging Perspective on Medico-Legal Autopsy: A Mini Review

“…Other potentially non-mixed, but easily containable low template DNA samples such as burned body or tissue which was subjected to a high temperature [46, [63][64][65], formalin-fixed samples [66][67][68], a piece of short hair with no root [69], or fingerprints of a touched surface and body parts [38,39,[70][71][72] often require special consideration. Depending on the complexity of various influences, and with lack of valid information concerning the initial environmental circumstances and the chemical and/or physical processes involved, the degree of damage of and/or quality of obtained DNA cannot be, at first, accurately predicted.…”