DOI: 10.1520/jfs14745j. Evaluation of an Alkaline Lysis Method for the Extraction of DNA from Whole Blood and Forensic Stains for STR Analysis

Evaluation of an Alkaline Lysis Method for the Extraction of DNA from Whole Blood and Forensic Stains for STR Analysis

10.1520/jfs14745j

Crossref journal-article

ASTM International

Journal of Forensic Sciences (381)

Abstract

Abstract A modified alkaline lysis protocol for extracting DNA from forensically relevant specimens is evaluated and compared with the chelex 100 method. For whole blood, bloodstains and sperm stains, both methods yielded comparable results after amplification for a pentameric STR locus (HumCD4). The main advantages of the new method are: only approximately ten minutes and two pipetting steps are necessary and the expenses for the extraction are extremely low as only NaOH, TrisHC1 buffer and a single microcentrifuge tube are required. Alkaline lysis also proved to yield DNA suitable for typing longer STRs by using dye-labeled primers and capillary electrophoresis. These advantages should render this protocol especially interesting for high-throughput laboratories in combination with multiplex PCR and fluorescent dye technology, although the storability of the extracts proved to be problematic.

Bibliography

Klintschar, M., & Neuhuber, F. (2000). Evaluation of an Alkaline Lysis Method for the Extraction of DNA from Whole Blood and Forensic Stains for STR Analysis. Journal of Forensic Sciences, 45(3), 669â673.

Authors 2

M Klintschar (first)
F Neuhuber (additional)

References 20 Referenced 56

10.1016/0098-2997(95)00003-Y / Molec Aspects Med / DNA technology in forensic applications by Hochmeister (1995)
10.1002/elps.1150190116 / Electrophoresis / Genotyping of forensic short tandem repeat (STR) systems based on sizing precision in a capillary electrophoresis instrument by Lazaruk (1998)
10.1111/j.1432-1033.1973.tb02881.x / Eur J Biochem / Isolation of high molecular weight DNA from mammalian cells by Gross-Bellard (1973)
10.1093/nar/16.3.1215 / Nucleic Acids Res / A simple salting out procedure for extracting DNA from human nucleated cells by Miller (1988)
{'key': '2025063017080927200_c5', 'first-page': '506', 'article-title': 'Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material', 'volume': '10', 'author': 'Walsh', 'year': '1991', 'journal-title': 'Biotechniques'} / Biotechniques / Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material by Walsh (1991)
10.1016/0005-2787(75)90283-X / Biochim Biophys Acta / Alkaline lysis of mammalian cells for sedimentation analysis of nuclear DNA. Conformation of released DNA as monitored by physical, electron microscopic and enzymological techniques by Parodi (1975)
10.1016/B978-0-12-281520-1.50013-5 / Protein folding by Ghélis (1982)
10.1006/abio.1993.1346 / Anal Biochem / A modified alkaline lysis method for the preparation of highly purified plasmid DNA from Escherichia coli by Felicello (1993)
10.1093/nar/7.6.1513 / Nucleic Acids Res / A rapid alkaline extraction procedure for screening recombinant plasmid DNA by Birnboim (1979)
10.1093/nar/21.17.4153 / Nucleic Acids Res / A simple method of preparing plant samples for PCR by Wang (1993)
{'key': '2025063017080927200_c11', 'first-page': '269', 'volume-title': 'A five minute procedure for extraction of genomic DNA from whole blood, semen and forensic stains for PCR', 'author': 'Dissing', 'year': '1996'} / A five minute procedure for extraction of genomic DNA from whole blood, semen and forensic stains for PCR by Dissing (1996)
10.1520/JFS14228J / J Forensic Sci / HumCD4-validation of a STR system for forensic purposes in an Austrian Caucasian population sample by Klintschar (1997)
10.1093/nar/20.19.5061 / Nucleic Acids Res / A rapid chemoluminescent method for quantitation of human DNA by Walsh (1992)
{'key': '2025063017080927200_c14', 'article-title': 'Population genetic studies on 9 tetrameric short tandem repeat loci using fluorescence dye labeled primers and capillary electrophoresis in the Egyptian population', 'author': 'Klintschar', 'year': '1999', 'journal-title': 'Forensic Sci Int'} / Forensic Sci Int / Population genetic studies on 9 tetrameric short tandem repeat loci using fluorescence dye labeled primers and capillary electrophoresis in the Egyptian population by Klintschar (1999)
10.1007/s001140050375 / Naturwissenschaften / DNA typing in forensic medicine and in criminal investigations: A current survey by Benecke (1997)
10.1385/0-89603-443-7:9 / Methods in Molecular Biology, Vol. 98: Forensic DNA profiling protocols / Recovery of DNA for PCR amplification from blood and forensic samples using a chelating resin by Willard (1998)
10.1520/JFS13711J / J Forensic Sci / DNA extraction strategies for amplified length polymorphism analysis by Comey (1994)
{'key': '2025063017080927200_c18', 'article-title': 'PCR inhibition and bloodstains', 'volume-title': 'Proceedings of the International Symposium on the forensic aspects of DNA analysis 189', 'author': 'Walsh', 'year': '1990'} / Proceedings of the International Symposium on the forensic aspects of DNA analysis 189 / PCR inhibition and bloodstains by Walsh (1990)
10.1520/JFS13607J / J Forensic Sci / Identification of the heme compound copurified with deoxyribonucleic acid (DNA) from bloodstains, a major inhibitor of polymerase chain reaction (PCR) amplification by Akane (1994)
10.1520/JFS12039J / J Forensic Sci / NaOH treatment to neutralize inhibitors of Taq polymerase by Bourke (1999)

Dates

Type	When
Created	10 years ago (Aug. 23, 2015, 8:27 a.m.)
Deposited	2 months ago (June 30, 2025, 5:08 p.m.)
Indexed	2 months ago (June 30, 2025, 5:40 p.m.)
Issued	25 years, 4 months ago (May 1, 2000)
Published	25 years, 4 months ago (May 1, 2000)
Published Print	25 years, 4 months ago (May 1, 2000)

Funders 0

None

BibTeX

@article{Klintschar_2000, title={Evaluation of an Alkaline Lysis Method for the Extraction of DNA from Whole Blood and Forensic Stains for STR Analysis}, volume={45}, ISSN={0022-1198}, url={http://dx.doi.org/10.1520/jfs14745j}, DOI={10.1520/jfs14745j}, number={3}, journal={Journal of Forensic Sciences}, publisher={ASTM International}, author={Klintschar, M and Neuhuber, F}, year={2000}, month=may, pages={669–673} }

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