Summary
A simple, practical approach for the extraction of PCR-amplifiable DNA for the HLA-DQa gene from bloodstains deposited on various substrates is described. DNA from bloodstains is purified using Chelex 100 ion-exchange resin and then amplified. If amplification is not achieved, the extract is washed through a Centricon 100 dialysis/concentration tube. If the second amplification of this extract produces a negative result, the extract is processed with Chelex 100 again. This approach has been found to be reliable, safe, efficient and economical.
Zusammenfassung
Eine einfache DNA-Extraktionsmethode zur direkten Amplifikation des HLA-DQa-Locus aus auf verschiedenen Spurenträgern angelegten Blutspuren wird beschrieben. Die Extraktion erfolgt mittels des Chelatbildners Chelex 100. Ist die Amplifikation inhibiert, wird das Extrakt in Centricon 100 Mikrokonzentrator-Röhrchen dialysiert. Bleibt auch eine zweite Amplifikation negativ, erfolgt eine erneute Reinigung mittels Chelex 100. Diese Vorgangsweise erwies sich als effizient, zuverlässig und praktikabel.
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References
Erlich HA (1989) PCR technology: principles and application for DNA amplification. Stockton Press, New York
Westwood SA, Werrett DJ (1990) An evaluation of the polymerase chain reaction method for forensic applications. Forensic Sci Int 45:201–215
Comey CT, Jung JM, Budowle B (1990) Validation studies on the analysis of the HLA-DOu locus using the polymerase chain reaction. J Forensic Sei (submitted)
Chelex protocols (1990): Amplitype™ User Guide. Cetus Corporation, Emeryville, CA
Paabo S, Gifford JA, Wilson AC (1988) Mitochondrial DNA sequences from a 7000-year old brain. Nucleic Acids Res 16:9775–9787
Budowle B, Waye JS, Shutler GG, Baechtel FS (1990)HaeIII — A suitable endonuclease for restriction fragment length polymorphism analysis of biological evidence samples. J Forensic Sci 35:530–536
Waye JS, Presley LA, Budowle B, Schutler GG, Fourney RM (1990) A simple and sensitive method for quantifying human genomic DNA in forensic specimen extracts. Biotechniques 7 852–855
Saiki RK, Walsh PS, Levenson CH, Erlich HA (1989) Genetic analysis of amplified DNA with immobilized sequence-specific oligonucleotide probes. Proc Natl Acad Sei USA 86:6230–6234
deFranchis R, Cross NCP, Foulkes NS, Cox TM (1988) A potent inhibitor of Taq polymerase copurifies with human genomie DNA. Nucleic Acids Res 16:10355
Walsh S, Blake E, Higuchi R (1989) PCR inhibition and bloodstains (abstract). Proceedings of the International Symposium on the Forensic Aspects of DNA Analysis. United States Government, Printing Office, Washington, DC
Chelex 100 and Chelex 20 chelating ion exchange resin instruction manual. Bio-Rad Laboratories, Richmond
.Singer-Sam J, Tanguay RL, Riggs A (1989) Use of Chelex to improve the PCR signal from a small number of cells. Amplifications 3:11
Mark HF, Bikales NM, Overberger CG, Menges G, Kroschwitz JI (1986) Encyclopedia of polymer science and engineering, vol 6. Wiley, New York, pp 632–645
Grayson M (1984) Encyclopedia of textiles, fibers and nonwoven fabrics. Wiley, New York
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Jung, J.M., Comey, C.T., Baer, D.B. et al. Extraction strategy for obtaining DNA from bloodstains for PCR amplification and typing of the HLA-DQa gene. Int J Leg Med 104, 145–148 (1991). https://doi.org/10.1007/BF01369719
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DOI: https://doi.org/10.1007/BF01369719