TY - JOUR
T1 - Identification of microsatellites from an extinct moa species using high-throughput (454) sequence data
AU - Allentoft, Morten E.
AU - Schuster, Stephan C.
AU - Holdaway, Richard N.
AU - Hale, Marie L.
AU - McLay, Emma
AU - Oskam, Charlotte
AU - Gilbert, M. Thomas P.
AU - Spencer, Peter
AU - Willerslev, Eske
AU - Bunce, Michael
PY - 2009/3
Y1 - 2009/3
N2 - Genetic variation in microsatellites is rarely examined in the field of ancient DNA (aDNA) due to the low quantity of nuclear DNA in the fossil record together with the lack of characterized nuclear markers in extinct species. 454 sequencing platforms provide a new high-throughput technology capable of generating up to 1 gigabases per run as short (200-400-bp) read lengths. 454 data were generated from the fossil bone of an extinct New Zealand moa (Aves: Dinornithiformes). We identified numerous short tandem repeat (STR) motifs, and here present the successful isolation and characterization of one polymorphic microsatellite (Moa-MS2). Primers designed to flank this locus amplified all three moa species tested here. The presented method proved to be a fast and efficient way of identifying microsatellite markers in ancient DNA templates and, depending on biomolecule preservation, has the potential of enabling high-resolution population genetic studies of extinct taxa. As sequence read lengths of the 454 platforms and its competitors (e.g., the SOLEXA and SOLiD platforms) increase, this approach will become increasingly powerful in identifying microsatellites in extinct (and extant) organisms, and will afford new opportunities to study past biodiversity and extinction processes.
AB - Genetic variation in microsatellites is rarely examined in the field of ancient DNA (aDNA) due to the low quantity of nuclear DNA in the fossil record together with the lack of characterized nuclear markers in extinct species. 454 sequencing platforms provide a new high-throughput technology capable of generating up to 1 gigabases per run as short (200-400-bp) read lengths. 454 data were generated from the fossil bone of an extinct New Zealand moa (Aves: Dinornithiformes). We identified numerous short tandem repeat (STR) motifs, and here present the successful isolation and characterization of one polymorphic microsatellite (Moa-MS2). Primers designed to flank this locus amplified all three moa species tested here. The presented method proved to be a fast and efficient way of identifying microsatellite markers in ancient DNA templates and, depending on biomolecule preservation, has the potential of enabling high-resolution population genetic studies of extinct taxa. As sequence read lengths of the 454 platforms and its competitors (e.g., the SOLEXA and SOLiD platforms) increase, this approach will become increasingly powerful in identifying microsatellites in extinct (and extant) organisms, and will afford new opportunities to study past biodiversity and extinction processes.
KW - Ancient DNA
KW - Extinct species
KW - High-throughput 454 sequencing
KW - Microsatellite development
UR - http://www.scopus.com/inward/record.url?scp=65449135172&partnerID=8YFLogxK
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U2 - 10.2144/000113086
DO - 10.2144/000113086
M3 - Article
C2 - 19317662
AN - SCOPUS:65449135172
SN - 0736-6205
VL - 46
SP - 195
EP - 200
JO - BioTechniques
JF - BioTechniques
IS - 3
ER -