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Frequent D-loop polymorphism in mtDNA enables genotyping of 1400-year-old human remains from Merowingian graves

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Zeller M., Mirghomizadeh M. F., Mirghomizadeh F., Wehner W. H.-D., Wehner H.-D., Blin B. N., Blin N.

Journal of Applied Genetics

2000

vol. 41

issue 4

285-292

Improvements of DNA extraction and amplification techniques presently enable DNA analysis of ancient DNA (aDNA) from samples which range from several hundred years of age up to possibly 5000 years. Taking advantage of the abundance of mitochondrial DNA and its polymorphic D-loop sequence, ten individuals from multiple burial sites of the Merowingian culture (South Germany), estimated to be about 1400 years old, were genotyped to determine possible kinship. Moreover, gonosomal DNA markers from the X- and Y-chromosome were applied for sex determination of the remains. In all individuals investigated, deviations from the Anderson mtDNA consensus sequence were observed, all representing substitutions (7 transitions and 3 transversions). Although such mutations have been reported from recent populations, our study constitutes the first description of these mtDNA mutations from numerous aDNA samples recovered from multiple burial sites. The results obtained by molecular anthropology can aid in describing kinship relations and burial customs of ancient remains.

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

84%

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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2 Journal of Applied Genetics

2 2000

2 Blin B. N.

2 Blin N.

1 Broghammer B. M.

1 Broghammer M.

1 Mirghomizadeh F.

1 Mirghomizadeh M. F.

1 Nicholson G. J.

1 Nicholson N. G. J.

1 Pusch C. M.

1 Scholz M.

1 Scholz S. M.

1 Wehner H.-D.

1 Wehner W. H.-D.

1 Zeller M.

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Frequent D-loop polymorphism in mtDNA enables genotyping of 1400-year-old human remains from Merowingian graves

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