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. 1998 Feb 1;26(3):857–859. doi: 10.1093/nar/26.3.857

Repair of degraded duplex DNA from prehistoric samples using Escherichia coli DNA polymerase I and T4 DNA ligase.

C M Pusch 1, I Giddings 1, M Scholz 1
PMCID: PMC147318  PMID: 9443981

Abstract

The most notable feature of DNA extracted from prehistoric material is that it is of poor quality. Amplification of PCR products from such DNA is consequently an exception. Here we present a simple method for the repair of degraded duplex DNA using the enzymes Escherichia coli DNA polymerase I and T4 DNA ligase. Adjacent sequences separated by nicks do not split up into intact strands during the denaturation step of PCR. Thus the target DNA is refractory to amplification. The proposed repair of nicked, fragmented ancient DNA results in an increase of amplification efficiency, such that the correct base order of the respective nuclear DNA segment can be obtained.

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Selected References

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  1. Hagelberg E., Clegg J. B. Isolation and characterization of DNA from archaeological bone. Proc Biol Sci. 1991 Apr 22;244(1309):45–50. doi: 10.1098/rspb.1991.0049. [DOI] [PubMed] [Google Scholar]
  2. Hänni C., Brousseau T., Laudet V., Stehelin D. Isopropanol precipitation removes PCR inhibitors from ancient bone extracts. Nucleic Acids Res. 1995 Mar 11;23(5):881–882. doi: 10.1093/nar/23.5.881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Hänni C., Brousseau T., Laudet V., Stehelin D. Isopropanol precipitation removes PCR inhibitors from ancient bone extracts. Nucleic Acids Res. 1995 Mar 11;23(5):881–882. doi: 10.1093/nar/23.5.881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Höss M., Jaruga P., Zastawny T. H., Dizdaroglu M., Päbo S. DNA damage and DNA sequence retrieval from ancient tissues. Nucleic Acids Res. 1996 Apr 1;24(7):1304–1307. doi: 10.1093/nar/24.7.1304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Krings M., Stone A., Schmitz R. W., Krainitzki H., Stoneking M., Päbo S. Neandertal DNA sequences and the origin of modern humans. Cell. 1997 Jul 11;90(1):19–30. doi: 10.1016/s0092-8674(00)80310-4. [DOI] [PubMed] [Google Scholar]
  6. Pusch C. A simple and fast procedure for high quality DNA isolation from gels using laundry detergent and inverted columns. Electrophoresis. 1997 Jun;18(7):1103–1104. doi: 10.1002/elps.1150180713. [DOI] [PubMed] [Google Scholar]
  7. Päbo S. Ancient DNA: extraction, characterization, molecular cloning, and enzymatic amplification. Proc Natl Acad Sci U S A. 1989 Mar;86(6):1939–1943. doi: 10.1073/pnas.86.6.1939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Päbo S., Higuchi R. G., Wilson A. C. Ancient DNA and the polymerase chain reaction. The emerging field of molecular archaeology. J Biol Chem. 1989 Jun 15;264(17):9709–9712. [PubMed] [Google Scholar]
  9. Taberlet P., Griffin S., Goossens B., Questiau S., Manceau V., Escaravage N., Waits L. P., Bouvet J. Reliable genotyping of samples with very low DNA quantities using PCR. Nucleic Acids Res. 1996 Aug 15;24(16):3189–3194. doi: 10.1093/nar/24.16.3189. [DOI] [PMC free article] [PubMed] [Google Scholar]

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