Metagenomics to Paleogenomics: Large-Scale Sequencing of Mammoth DNA

От метагеномики к палеогеномике: крупномасштабное секвенирование ДНК мамонта
Hendrik N. Poinar, C. Schwarz, Ji Qi, Beth Shapiro, R. D. E. MacPhee, Bernard Buigues, Alexei Tikhonov, Daniel H. Huson, Lynn P. Tomsho, Alexander F. Auch, Markus Rampp, Webb Miller, Stephan C. Schuster
2005-12-20

emulsion polymerase chain reactionmetagenomic sequencingpaleogenomicspyrosequencingwoolly mammoth DNA
We sequenced 28 million base pairs of DNA in a metagenomics approach, using a woolly mammoth (Mammuthus primigenius) sample from Siberia. As a result of exceptional sample preservation and the use of a recently developed emulsion polymerase chain reaction and pyrosequencing technique, 13 million base pairs (45.4%) of the sequencing reads were identified as mammoth DNA. Sequence identity between our data and African elephant (Loxodonta africana) was 98.55%, consistent with a paleontologically based divergence date of 5 to 6 million years. The sample includes a surprisingly small diversity of environmental DNAs. The high percentage of endogenous DNA recoverable from this single mammoth would allow for completion of its genome, unleashing the field of paleogenomics.
1
A metagenomic approach generated 28 million base pairs of sequence from a Siberian woolly mammoth sample.
2
Exceptional preservation and emulsion PCR–pyrosequencing yielded 13 million base pairs of identifiable mammoth DNA, representing 45.4% of reads.
3
Mammoth sequences showed 98.55% identity to African elephant DNA, supporting a divergence time of 5–6 million years.
4
The high endogenous DNA yield from one mammoth sample suggests that completing the mammoth genome is feasible and enables paleogenomic research.
5
The sample contained unexpectedly low diversity of environmental DNA, facilitating recovery of endogenous mammoth sequences.

A woolly mammoth (Mammuthus primigenius) DNA sample from Siberia

The recoverability, taxonomic identification, and evolutionary divergence of endogenous mammoth DNA, including its potential for genome reconstruction

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2005-12-20
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Authors
Hendrik N. Poinar
C. Schwarz
Ji Qi
Beth Shapiro
R. D. E. MacPhee
Bernard Buigues
Alexei Tikhonov
Daniel H. Huson
Lynn P. Tomsho
Alexander F. Auch
Markus Rampp
Webb Miller
Stephan C. Schuster
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