A high volume extraction and purification method for recovering DNA from human bone

Marshall, Pamela L.; Stoljarova, Monika; Schmedes, Sarah E.; King, Jonathan L.; Budowle, Bruce

doi:10.1016/j.fsigen.2014.06.011

Cited by 28 publications

(18 citation statements)

References 19 publications

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“…Every step involving ancient samples was carried out in laboratories exclusively dedicated to ancient DNA analysis. DNA extraction was performed by adsorption to silica gel membrane, following a protocol modified from Marshall et al 17 Hypervariable segments HVS-I and HVS-II of the mtDNA control region were amplified by PCR using five overlapping fragments of ∼ 175 base pairs (bp) each one (Supplementary Table S1). Samples were subjected to different additional analyses to confirm 1 the results and to probe their authenticity, such as cloning amplified sequences, analysing haplogroup diagnostic polymorphisms located in the mtDNA coding region by different techniques, and the independent analysis of some samples by an external laboratory of Strasbourg (France).…”

Section: Methodsmentioning

confidence: 99%

Ancient mitochondrial lineages support the prehistoric maternal root of Basques in Northern Iberian Peninsula

et al. 2017

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The Basque population inhabits the Franco-Cantabrian region in southwest Europe where Palaeolithic human groups took refuge during the Last Glacial Maximum. Basques have been an isolated population, largely considered as one of the most ancient European populations and it is possible that they maintained some pre-Neolithic genetic characteristics. This work shows the results of mitochondrial DNA analysis of seven ancient human remains from the Cave of Santimamiñe in the Basque Country dated from Mesolithic to the Late Roman period. In addition, we compared these data with those obtained from a modern sample of Basque population, 158 individuals that nowadays inhabits next to the cave. The results support the hypothesis that Iberians might have been less affected by the Neolithic mitochondrial lineages carried from the Near East than populations of Central Europe and revealed the unexpected presence of prehistoric maternal lineages such as U5a2a and U3a in the Basque region. Comparison between ancient and current population samples upholds the hypothesis of continuity of the maternal lineages in the area of the Franco-Cantabrian region.

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Section: Methodsmentioning

confidence: 99%

Ancient mitochondrial lineages support the prehistoric maternal root of Basques in Northern Iberian Peninsula

et al. 2017

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“…Samples were then grounded into a fine powder in a 6750 Freezer Mill (Spex SamplePrep, USA) under liquid nitrogen to avoid DNA degradation. Afterwards, about 500 mg of powder was used for a silica-based DNA extraction method (Qiagen, Hilden, Germany) (modified of [ 4 ]) adapted to the Hi-Flow ® DNA Purification Spin Columns (Generon, Berkshire, UK). A final concentration step to 30–40 μl was carried out using Amicon-30 columns (Millipore, MA, USA).…”

Section: Methodsmentioning

confidence: 99%

Mitochondrial DNA Reveals the Trace of the Ancient Settlers of a Violently Devastated Late Bronze and Iron Ages Village

et al. 2016

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La Hoya (Alava, Basque Country) was one of the most important villages of the Late Bronze and Iron Ages of the north of the Iberian Peninsula, until it was violently devastated around the 4th century and abandoned in the 3rd century B.C. Archaeological evidences suggest that descendants from La Hoya placed their new settlement in a nearby hill, which gave rise to the current village of Laguardia. In this study, we have traced the genetic imprints of the extinct inhabitants of La Hoya through the analysis of maternal lineages. In particular, we have analyzed the mitochondrial DNA (mtDNA) control region of 41 human remains recovered from the archaeological site for comparison with a sample of 51 individuals from the geographically close present-day population of Laguardia, as well as 56 individuals of the general population of the province of Alava, where the archaeological site and Laguardia village are located. MtDNA haplotypes were successfully obtained in 25 out of 41 ancient samples, and 14 different haplotypes were identified. The major mtDNA subhaplogroups observed in La Hoya were H1, H3, J1 and U5, which show a distinctive frequency pattern in the autochthonous populations of the north of the Iberian Peninsula. Approximate Bayesian Computation analysis was performed to test the most likely model for the local demographic history. The results did not sustain a genealogical continuity between Laguardia and La Hoya at the haplotype level, although factors such as sampling effects, recent admixture events, and genetic bottlenecks need to be considered. Likewise, the highly similar subhaplogroup composition detected between La Hoya and Laguardia and Alava populations do not allow us to reject a maternal genetic continuity in the human groups of the area since at least the Iron Age to present times. Broader analyses, based on a larger collection of samples and genetic markers, would be required to study fine-scale population events in these human groups.

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“…For semen and other body fluid mixtures (blood/semen and saliva/semen mixtures), the differential extraction method according to Giusti et al [22] was used to extract DNA. Bone samples were extracted as described by Marshall et al [23]. Quantifiler Human DNA Quantification Kit (Thermo Fisher, Foster City, CA, USA) on an ABI Prism 7500 Sequence Detection system (Thermo Fisher) was used to estimate the quantity of extracted DNA [24].…”

Section: Sample Preparationmentioning

confidence: 99%

An evaluation of the PowerSeq™ Auto System: A multiplex short tandem repeat marker kit compatible with massively parallel sequencing

Zeng

King

Hermanson

et al. 2015

Forensic Science International: Genetics

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A high volume extraction and purification method for recovering DNA from human bone

Cited by 28 publications

References 19 publications

Ancient mitochondrial lineages support the prehistoric maternal root of Basques in Northern Iberian Peninsula

Ancient mitochondrial lineages support the prehistoric maternal root of Basques in Northern Iberian Peninsula

Mitochondrial DNA Reveals the Trace of the Ancient Settlers of a Violently Devastated Late Bronze and Iron Ages Village

An evaluation of the PowerSeq™ Auto System: A multiplex short tandem repeat marker kit compatible with massively parallel sequencing

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