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Molecular phylogenetics employing modern and ancient DNA

100%

Pusch C. M., Broghammer M., Blin N.

Journal of Applied Genetics

2003

vol. 44

issue 3

269-290

Comparative studies of DNA in recent populations and characterisation of ancient hereditary material have contributed very interesting facts to our understanding of evolution of modern mankind. Analysis of DNA homology in related species, assessment of mutations and polymorphisms in various populations and new DNA sequence data from prehistoric finds allowed ? via sophisticated DNA extraction techniques, PCR, sequencing and digitalised processing of genetic information ? insights into possible roots of Homo sapiens and related species, migration patterns and ancient cultural habits, thus enriching the palaeoanthropological discipline. However, a presentation of this development would not be complete without pointing towards the methodological limitations and manifold presentations burdened with artifacts, data misinterpretation and unjustified conclusions. Presently, this modern field of research is in its consolidation phase and new parameters for quality control and authentication are being implemented to avoid spectacular but unfounded reports. It is expected that most of the problems connected to old biomolecules may be closely related to fossilisation parameters. The future challenge will be the full understanding of the complex and multi-faceted processes underlying diagenesis, including the elucidation of nucleic acid ?postmortem damage?.

Adaptor-mediated amplification of minute amounts of severely fragmented ancient nucleic acids

64%

Pusch C. M., Blin B. N., Blin N., Broghammer B. M., Broghammer M., Nicholson N. G. J., Nicholson G. J., Scholz S. M., Scholz M.

Journal of Applied Genetics

2000

vol. 41

issue 4

303-315

It has been repeatedly shown that high copy number mitochondrial DNA sequences can be recovered from ancient samples. A significant increase in the volume of information available to researchers will be observed when the amplification of nuclear DNA becomes commonplace and reproducible. To this end we established a modification of the Rapid Amplification of cDNA Ends (RACE) procedure normally used for the generation of cDNA ends from adaptor-ligated expressed sequence tag libraries. The modifications were designed to specifically address the problems associated with the highly damaged nucleic acids extracted from palaeontological specimens. For this study we used 6 human samples dating to 450 AD and ~6.500 BP that were refractory to reliable amplification of single copy loci by PCR. Racemate contents (ratio of D/L enantiomers) of aspartic acid, alanine, and leucine also indicated that no amplifiable DNA is present in 5 of the 6 samples. The proposed technique allowed us (i) to amplify four X-chromosomal loci from 5 human specimens, and (ii) to correct allelic drop-out phenomena at the amelogenin locus in one individual, thus showing that the threshold of 80 ? 10?3 for D/Lasp as a borderline for the presence/absence of amplifiable aDNA requires reassessment. Reliability of the proposed technique (i.e. amplification of DNA sequences endogenous to the find) was validated by the application of ?ancient RACE? (aRACE) to prehistoric animal samples.

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Molecular phylogenetics employing modern and ancient DNA

Adaptor-mediated amplification of minute amounts of severely fragmented ancient nucleic acids