Bibliography and abstracts on DNA isolation from deceased tissues.

Genelex has developed specific methodologies that greatly enhance our ability to recover usable DNA. We have demonstrated this ability with bones, teeth, decomposed tissues, and a wide variety of archived and chemically preserved specimens. The follwing abstracts offer additional information on isolating DNA from various postmortem samples..

J: Forensic Sci 1993 May;38(3):686-90 Related Articles, Books, LinkOut

DNA fingerprinting from tissues after variable postmortem periods.

Ludes B, Pfitzinger H, Mangin P

Institut de Medecine Legale, Strasbourg, France.

DNA typing is a useful tool in forensic cases for determining the identity of remains of humans who have been dead for various periods of time. DNA fingerprinting can be achieved only if high molecular weight DNA (HMWDNA) is extracted from the tissue samples of the bodies even after a long postmortem delay. Analyses were performed on various tissues collected during forensic autopsies of 24 bodies known postmortem ages.Tissues such as blood and kidney were found to be unsuitable for DNA fingerprinting because of a rapid degradation of the DNA after a period of one week. HMWDNA could be successfully extracted from brain cortex regardless of postmortem age.

J: Forensic Sci 1997 Jul;42(4):708-14 Related Articles, Books

The applicability of formalin-fixed and formalin fixed paraffin embedded tissues in forensic DNA analysis.

Romero RL, Juston AC, Ballantyne J, Henry BE

Washoe County Sheriff's Office, Forensic Science Division, Reno, NV, USA.

Historically, formalin fixed (FF) tissues could not be used as a source of DNA in forensic science due to the fact that the DNA was too degraded for DNA analysis. With the introduction of the polymerase chain reaction (PCR) technique to forensic science, the usefulness of DNA from this biological material has been re-evaluated. This study evaluates the potential use of DNA from FF and formalin fixed paraffin embedded (FFPE) tissues in 13 PCR systems; HLA DQ alpha, LDLR, GYPA, HBGG, D7S8, GC, D1S80, vWA31, THO1, F13A1, FES/FPS, TPOX, and CSF1PO. The first six, HLA DQ alpha, LDLR, GYPA, HBGG, D7S8, and GC are reverse dot blot systems, D1S80 is an amplified fragment length polymorphism (AmpFlp) system and the others are short tandem repeats (STRs). This study shows that FFPE tissue which has not been fixed in formalin for more than three days is a useful source of DNA for 12 of the 13 PCR systems. In contrast, FF tissue did not prove to be a reliable source of DNA for the PCR techniques examined here.

Nature 1991 Aug 1;352(6334):427-9 Related Articles, Books

Identification of the skeletal remains of a murder victim by DNA analysis.

Hagelberg E, Gray IC, Jeffreys AJ MRC

Molecular Haematology Unit, University of Oxford, John Radcliffe Hospital, UK.

There is considerable anthropological and forensic interest in the possibility of DNA typing skeletal remains. Trace amounts of DNA can be recovered even from 5,500-year-old bones and multicopy human mitochondrial DNA sequences can frequently be amplified from such DNA using the polymerase chain reaction (PCR). But given the sensitivity of PCR, it is very difficult to exclude contaminating material. We now report the successful identification of the 8-year-old skeletal remains of a murder victim, by comparative typing of nuclear microsatellite markers in the remains and in the presumptive parents of the victim. This analysis establishes the authenticity of the bone DNA and the feasibility of bone DNA typing in forensic investigations.

Comments:

Comment in: Nature 1991 Aug 1;352(6334):381-2

Comment in: Nature 1991 Nov 14;354(6349):113

Comment in: Nature 1992 Apr 9;356(6369):471

J Forensic Sci 1988 Jan;33(1):144-53 Related Articles, Books, LinkOut

The autodegradation of deoxyribonucleic acid (DNA) in human rib bone and its relationship to the time interval since death.

Perry WL 3d, Bass WM, Riggsby WS,

Sirotkin K College Scholars Program, University of Tennessee, Knoxville. This research explored the feasibility of using the degradation rate of deoxyribonucleic acid (DNA) in human rib bone to determine the time interval since death. Postmortem human rib samples were surface sterilized and incubated under sterile conditions in either high or low humidity conditions at room temperature for a period of weeks. At selected times, portions of the bone were cut away, and the DNA from these samples was extracted and subjected to strand separating gel electrophoresis. The DNAs in the gels were transferred to a nylon membrane, preserving their relative positions as in the gel, and probed with radioactive total genomic human DNA. Autoradiograms produced were scanned and digitized. When the samples were incubated under identical conditions, the degradation rate of DNA in samples from different individuals appeared very similar. The DNA degradation rate may vary with temperature and humidity more than it varies between individuals.

Cancer Res 1986 Jun;

46(6):2964-9 Related Articles, Books

Southern blot analysis of DNA extracted from formalin-fixed pathology specimens.

Dubeau L, Chandler LA, Gralow JR, Nichols PW, Jones PA

We have developed a method for the extraction of DNA from formalin-fixed, paraffin-embedded pathology specimens. High-molecular-weight DNA was recovered from well-fixed nonautolyzed samples of viable tissue. DNA recovered from samples exposed to picric acid or mercuric chloride containing fixatives was not intact. Increasing the formalin fixation time decreased the amount of intact DNA available. When these limitations were taken into consideration, the procedure allowed for the removal of degraded and chemically modified DNA from the preparation, and the final product was suitable for quantitative and qualitative analysis by Southern or dot blotting techniques. Digestion with methylation-sensitive restriction endonucleases showed that DNA methylation patterns were not altered after formalin fixation.

J Forensic Sci 1997 Nov;42(6):1126-35 Related Articles, Books

Comparison of three DNA extraction methods on bone and blood stains up to 43 years old and amplification of three different gene sequences.

Cattaneo C, Craig OE, James NT,

Sokol RJ Istituto di Medicina Legale e delle Assicurazioni, Universita degli Studi di Milano, Italy.

Extraction of amplifiable DNA-from degraded human material in the forensic context remains a problem, and maximization of yield and elimination of inhibitors of the Polymerase Chain Reaction (PCR) are important issues which rarely feature in comparative studies. The present work used PCR amplification of three DNA sequences (HLA DPB1, amelogenin and mitochondrial) to assess the efficiency of three methods for extracting DNA (sodium acetate, magnetic beads and glass-milk) from 32 skeletal samples and 25 blood stains up to 43 years old. The results, analyzed using multivariate statistics, confirmed that the extraction method was crucial to the subsequent detection of amplification products; the glass-milk protocol performed better than sodium acetate, which was better than magnetic beads. Successful amplification also depended on gene sequence, multiple copy mitochondrial sequences performing best; however, with the singly copy sequences, the longer HLA DPB1 (327 bp) being detected just as often as the shorter amelogenin (106/112 bp). Amplification products were obtained more frequently from blood stains than bone, perhaps reflecting differences inherent in the material, and from younger compared with older specimens, though plateauing seemed to occur after 10 years. PCR inhibitors were more frequent in sodium acetate extracts.