Report

An individual with Sarmatian-related ancestry in

Roman Britain
Graphical abstract Authors
Marina Silva, Thomas Booth,
Joanna Moore, ..., David Bowsher,
Janet Montgomery, Pontus Skoglund

Correspondence
marina.silva@crick.ac.uk (M.S.),
janet.montgomery@durham.ac.uk (J.M.),
pontus.skoglund@crick.ac.uk (P.S.)

In brief
Silva et al. identify a Roman-era individual
buried in rural Britain with ancestry
related to the Caucasus- and Sarmatian-
associated groups. Isotope analysis
evidences long-range childhood mobility.
These results may be linked to historical
movements of Sarmatians and highlight
long-distance mobility reaching rural
regions of the Roman Empire.

Highlights
d Ancestry outlier identified in rural Roman Britain dating to
126–228 cal. CE

d Genetically related to contemporary Sarmatian- and

Caucasus-associated groups

d Stable isotope analysis reveals life history of mobility

d Deployment of Sarmatian cavalry to Britain in 175 CE is a

plausible explanation

Silva et al., 2024, Current Biology 34, 204–212

January 8, 2024 ª 2023 The Author(s). Published by Elsevier Inc.
https://doi.org/10.1016/j.cub.2023.11.049 ll
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An individual with Sarmatian-related
ancestry in Roman Britain
Marina Silva,1,* Thomas Booth,1 Joanna Moore,2 Kyriaki Anastasiadou,1 Don Walker,3 Alexandre Gilardet,1
Christopher Barrington,4 Monica Kelly,1 Mia Williams,1 Michael Henderson,3 Alex Smith,5 David Bowsher,3
Janet Montgomery,2,* and Pontus Skoglund1,6,*
1Ancient Genomics Laboratory, The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK
2Department of Archaeology, Durham University, Lower Mountjoy, South Rd, DH1 3LE, Durham, United Kingdom
3Museum of London Archaeology (MOLA), Mortimer Wheeler House, 46 Eagle Wharf Road, London N1 7ED, UK
4Bioinformatics and Biostatistics, The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK
5Headland Archaeology, 13 Jane Street, Edinburgh EH6 5HE, UK
6Lead contact

*Correspondence: marina.silva@crick.ac.uk (M.S.), janet.montgomery@durham.ac.uk (J.M.), pontus.skoglund@crick.ac.uk (P.S.)

https://doi.org/10.1016/j.cub.2023.11.049

SUMMARY

In the second century CE the Roman Empire had increasing contact with Sarmatians, nomadic Iranian
speakers occupying an area stretching from the Pontic-Caspian steppe to the Carpathian mountains, both
in the Caucasus and in the Danubian borders of the empire.1–3 In 175 CE, following their defeat in the Marco-
mannic Wars, emperor Marcus Aurelius drafted Sarmatian cavalry into Roman legions and deployed 5,500
Sarmatian soldiers to Britain, as recorded by contemporary historian Cassius Dio.4,5 Little is known about
where the Sarmatian cavalry were stationed, and no individuals connected with this historically attested
event have been identified to date, leaving its impact on Britain largely unknown. Here we document Cauca-
sus- and Sarmatian-related ancestry in the whole genome of a Roman-period individual (126–228 calibrated
[cal.] CE)—an outlier without traceable ancestry related to local populations in Britain—recovered from a
farmstead site in present-day Cambridgeshire, UK. Stable isotopes support a life history of mobility during
childhood. Although several scenarios are possible, the historical deployment of Sarmatians to Britain pro-
vides a parsimonious explanation for this individual’s extraordinary life history. Regardless of the factors
behind his migrations, these results highlight how long-range mobility facilitated by the Roman Empire
impacted provincial locations outside of urban centers.

RESULTS In a principal-component analysis (PCA), Offord Cluny 203645

is differentiated from all other sampled Roman individuals from
An ancestry outlier in rural Roman Cambridgeshire Britain, excavated from a Roman cemetery at Driffield Terrace,
Human remains were recovered from an isolated burial during in the present-day city of York, northeast England (England_
excavations near the village of Offord Cluny led by MHI (Museum Roman, excluding a previously described outlier with ancestry
of London Archaeology [MOLA] Headland Infrastructure) in related to Near Eastern populations).9 Instead, Offord Cluny
advance of the National Highways A14 road development in 203645 is most similar to present-day individuals from Anatolia
Cambridgeshire, England (Figures 1A and S1A). We generated and the Caucasus (Figures 1C and S2A). Specifically, he shows
a
5.43 whole genome from the cochlea portion of the temporal affinities to Late Bronze Age individuals from Armenia (Armenia_
bone of the Offord Cluny skeleton (Sk 203645, Burial 20.507, LBA) and individuals recovered from Alan-associated contexts in
C10271), using single-stranded DNA library preparation (STAR the North Caucasus (Russia_Sarmatian_Alan, dating to 450–
Methods; Data S1A). A tooth was directly radiocarbon dated to 1350 CE,10 generally considered as part of the Sarmatian
126–228 cal. CE (95% confidence, SUERC-105720), in the confederation11), but not with individuals from Armenia who
early-mid Roman period (Figure 1B). The skeleton was only post-date the Bronze Age (here defined as Armenia_Antiquity12)
moderately well preserved macroscopically: although osteolog- (Figure 1D).
ical analysis of the remains suggested the individual was 18–25 Similarly, direct statistical tests in the form of f4-statistics
years old, it was not possible to produce a sex estimate. consistently show that the genetic ancestry of the Offord
Although there were some osteological indications of minor Cluny individual was different from the ancestry of Romano-
trauma in the past, there was nothing to suggest a cause of British individuals from Driffield Terrace, and he instead
death. Assessment of karyotypic sex6 using the sequenced shared genetic affinities with ancient populations from the
genome established that the remains belonged to a male individ- Caucasus and Pontic-Caspian region (Figure S3 and
ual (XY). Data S2A).

204 Current Biology 34, 204–212, January 8, 2024 ª 2023 The Author(s). Published by Elsevier Inc.
This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
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A C

Figure 1. Ancestry outlier Offord Cluny 203645

(A) Map of the site, showing the excavation area and the location of burial relative to Roman roads and Roman Godmanchester. Burial shown in Figure S1; for
sequencing metrics and uniparental haplogroups see Data S1.
(B) Calibrated radiocarbon date (126–228 cal. CE) of Offord Cluny 203645’s second right maxillary molar using OxCal v4.47 and IntCal208 (1,867 ± 16 years before
present [BP], SUERC-105720 [GU61561]).
(C) Principal component analysis (PCA) showing Offord Cluny 203645 (yellow square) and other previously published ancient individuals projected onto PCs
defined by 1,388 present-day western Eurasian individuals from the Affymetrix Human Origins (HO)
600k SNP panel. Individuals included in the populations
used as sources in the qpWave/qpAdm models are highlighted, with additional individuals from the same regions colored according to geography (shown in D).
For a detailed caption of all projected ancient individuals see Figure S2A; f4-statistics shown in Figure S3. Present-day individuals are indicated by the first 3 letters
of their population label, as reported in Data S2E.
(D) Map of ancient individuals included in PCA (with added jitter) and approximate calibrated dates of populations used as references in proximal models tested
with qpWave/qpAdm framework (shown in Figure 2). Offord Cluny 203645 is represented by a yellow square. Data points colored according to geography and
data type (whole-genome shotgun sequencing or ‘‘1240k’’ SNP capture), additional individuals are color-coded in Figure S2A.
See also Data S1 and S2.

Analysis of the Y chromosome and mitochondrial DNA (a sub-branch of R1b1a1b1b/R1b-Z2103) (Data S1B). This line-
(mtDNA) of Offord Cluny 203645, tracing paternal and maternal age has been previously identified in skeletal remains ranging
lineages, respectively, also point to ancestry from outside of from the Late Bronze Age to the Urartian period recovered
western Europe, in particular his paternal lineage: R1b-Y13369 from present-day Armenia,13 whereas its present-day phylogeny

Current Biology 34, 204–212, January 8, 2024 205

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A B

Figure 2. Ancestry modeling for Offord Cluny 203645

(A) Rotating models tested using qpWave/qpAdm framework. Armenia_LBA was excluded when testing 2-source models (shown in C). Models shown in (B) and
(C) used England_Roman; models using England_IA are shown in Data S2B and S2C.
(B) Location of populations included in the qpWave model (South_Africa_400BP and Yana_UP not shown) and p value for the single-source model accepted
(p value > 0.05); additional tested models shown in Data S2B. Location of the A14 site where Offord Cluny 203645 was found is indicated by a yellow square.
(C) Accepted 2-source qpAdm model for individual Offord Cluny 203645 when rotating through temporally proximal source (p > 0.05); all tested models shown in
Data S2C. Models using distal sources shown in Figure S2.
See also Data S2.

is dominated by samples from the Caucasus, Anatolia, and Near 203645.12,13 Therefore, Armenia_LBA is likely not a good repre-
East (Yfull tree v.11.01.00). Offord Cluny carried mtDNA hap- sentative of the ancestry observed in the Caucasus in the first
logroup K1a (Data S1C), found in Pre-Pottery Neolithic Anatolia millennium CE (Figure 1C). With this in mind, we tested additional
and the Levant, and in Europe since the Neolithic.14,15 Although models excluding Armenia_LBA (Figure 2C), which were consis-
subclades of haplogroup K1a, found at frequencies of
5% tent with Offord Cluny 203645 carrying
24%–34% of his
across all regions in the UK Biobank dataset,16 have been previ- ancestry from a source close to Sarmatian groups from the
ously identified in ancient individuals from Britain ranging from Pontic-Caspian region (either Russia_Sarmatian_PonticSteppe
the Neolithic to the early Medieval period, these all belong to or Russia_Sarmatian_SouthernUrals), in addition to ancestry
different sublineages than the one observed in Offord Cluny.17–21 from a source most similar to Armenia_Antiquity (p values
ranging from 0.062 to 0.124, and standard errors (SEs) varying
Relationship to Caucasus and Sarmatian groups from 5% to 6%, depending on the model; Data S2C). We
With the PCA having established the broad affinities of Offord note that a third similar model, with Russia_Sarmatian_Alan
Cluny 203645, we moved on to testing explicit ancestry models and Armenia_Antiquity as sources, is just under the threshold
with the qpWave/qpAdm framework. This approach allows us to of significance (p value
0.030, Data S2C). Overall, our results
test ancestry models and statistically reject those that do not fit suggest that there may have been substantial diversity among
the data. Our goal was to find models that uniquely fit the groups identified as Sarmatians, some of which could have
ancestry of the Offord Cluny 203645 individual—i.e., where all had ancestry that in our data is most closely represented by
other models of similar complexity (number of distinguishable Armenia_Antiquity.
ancestries) are rejected—with the caveat that we are limited to
the data available in the literature from approximately contempo- Stable isotopes support long-distance mobility
raneous periods from other regions. We first tested different sin- The results of the carbon (C), nitrogen (N), oxygen (O), and stron-
gle-source qpWave models rotating through different popula- tium (Sr) isotope analyses are presented in Figure 3. The
87
tions (STAR Methods), with a focus primarily on populations Sr/86Sr value from Offord Cluny 203645’s second mandibular
from the Caucasus and the Pontic-Caspian steppe, in addition molar (reflecting the first 5 to 6 years of his childhood22) was
to other populations from south and northern Europe (Figure 2A). 0.709037 ± 0.000012 (2 SE), and strontium concentration from
The only accepted single source is Armenia_LBA (p values = the same tooth was 104.2 parts per million (ppm), both of which
0.345 and 0.560), whereas Armenia_Antiquity, Sarmatian are within the range expected for Britain23,24 (Figure 3A and Data
groups, and populations from Britain (England_Roman or Eng- S3A). However, this is a common 87Sr/86Sr ratio that can be pro-
land_IA) are rejected as single sources (Figure 2B and Data S2B). duced by a wide range of geological terrains, and humans with
However, Armenia_LBA dates to
1200–850 BCE and thus similar values can be found in a variety of places. On the other
predates Offord Cluny 203645 by up to approximately one mil- hand, d18O values were lower than what would be expected if
lennium. Recent studies revealed ancestry changes in Armenia he had spent the first years of his childhood in Britain (Figure 3A
during the first millennium BCE, which resulted in different and Data S3A) and are instead indicative of regions with a colder
ancestry patterns in the region by the time of Offord Cluny or more continental climate, being consistent with levels of

206 Current Biology 34, 204–212, January 8, 2024

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A British
ritish oxygen range old, which could reflect at least two periods of movement across
0.7106 West Europe within the first
14 years of his life. It is not possible to
East
distinguish a gradual one-way transition in diet over several
0.7101
years of life from a fairly rapid change, due to increased overlap-
0.7096
ping in the orientation of the dentine incremental layers.31,32
Nevertheless, the gradual drop in d13C values observed after
Sr/86Sr

Seawater
0.7091 the age of 9 could reflect either a sustained increased consump-
Cambridgeshire
Cambridg
geshire
87

Offord
tion of C3 crops over several years or possibly a multi-year migra-
0.7086
tion, e.g., westward across Europe to Britain, through regions of
0.7081 gradually diminishing availability of C4 foods such as millet.

0.7076
15 16 17 18 19 DISCUSSION
δ18OVSMOW ‰
B -11 13
We have shown that the ancestry of Offord Cluny 203645 did not
-12 12 match that of the overall Romano-British population and that,
-13
instead, he shared genetic affinities with groups from the Cauca-
11
sus and the Pontic-Caspian steppe. Complex patterns of
-14
ancestry in the Caucasus12,13 and sparse sampling in the region,
δ13CVPDB ‰

10
δ15NAIR ‰

-15 particularly in the North Caucasus, covering the first four cen-
9 turies CE hinder the identification of a single proximal source
-16
for his ancestry. Future sampling in western Eurasia—and spe-
8
-17 cifically in the Pontic region and/or the North Caucasus—
7 covering the first and second centuries CE will have the potential
-18
to help narrow down Offord Cluny 203645’s ancestry, possibly
-19 6 allowing the identification of a single temporally proximal source
of ancestry.
-20 5
2 3 4 5 6 7 8 9 10 11 12 13 14 Genetics alone provide little insight on mobility within the life-
Approximate age (years)
time of one individual. Isotopic information is necessary for
investigating lifetime mobility patterns. Taken together, the C,
Figure 3. Stable isotope analyses N, Sr, and O isotope analyses indicate that Offord Cluny
(A) Offord Cluny 203645 human tooth enamel strontium (Sr) and oxygen
203645 spent the first 5 to 6 years of his childhood in a more
(O) isotope data (Data S3A) alongside mean (± 1 SD) regional comparative
data.28,29 The horizontal dotted lines represent the bioavailable Sr isotope eastern and arid continental location. This could include regions
range for Cambridgeshire.23 The shaded green and yellow boxes represent the within the empire, such as the northeastern Alps, but also areas
2 SD O isotope range expected for east and west Britain, respectively.24 beyond its borders, such as the mountainous regions of the Car-
Analytical error for O is 0.28&, 1 SD, and Sr is within the symbol. pathians or the Greater Caucasus. The incremental C and N sta-
(B) Diet changes in the first 14 years of Offord Cluny 203645’s life as indicated ble isotope analysis provided detailed information into Offord
by incremental dentine d13C and d15N data (second right mandibular molar,
Cluny 203645’s complex life history of long-distance migration,
M2) plotted against approximate age in years (see also Data S3B).
revealing two moments of dietary change: first at
5 years of
age, from a predominately C4 to a mixed C3/C4 diet, and then
precipitation recorded today in regions at high altitude.25 Similar again at
9 years of age to a diet based predominantly on C3 re-
combinations of Sr and O isotope ratios have been observed in sources, possibly reflecting two episodes of migration (Figure 3).
Roman-period populations in continental Europe.26,27 Linear defects, or enamel hypoplasia, on the crowns of nine teeth
Offord Cluny 203645 had high d13C values combined with low from Offord Cluny 203645 may reflect periods of arrested growth
d15N values, indicating a childhood diet rich in non-native C4 during episodes of malnutrition or illness.33,34 The location of
crops with little input from marine resources. Incremental these defects suggests they occurred around the age of 5 years,
dentine analysis (Figure 3B and Data S3B) revealed that his overlapping with the timing of the first observed shift in diet, and
diet underwent a substantial change around the age of 5 years, might therefore reflect physiological stress associated with die-
when d13C values drop from
12& to
16&, reflecting a tary changes and possible migration. The two shifts in diet might
clear shift from eating predominantly C4 plant protein to eating reflect a hiatus in his journey westwards before reaching Britain
a mixed C3/C4 diet with a possible increase in meat protein indi- and would be consistent with a period of time spent in central or
cated by a concomitant rise in d15N. A second change in diet southeastern Europe. The d13C value corresponding to
13
occurred after the age of 9, when the d13C profile started falling, years of age is closer to (but still slightly more elevated than)
reaching
19& around the age of 13, which is approaching an the values typically observed in Roman Britain,35,36 and thus it
entirely C3 based diet. As there is no clear evidence of wide con- is possible that he only moved to Britain later in his life.
sumption of C4 crops during the Roman occupation of Britain The impact of (possibly transient) long-distance individual
(despite some sporadic findings of millet)30 and they were not mobility and admixture at urban sites during historical pe-
common components of diet in western provinces of the Roman riods37,38 has been recently highlighted across a variety of sites
Empire, these two shifts in diet could represent a relocation in Europe, North Africa, the Caucasus, and the Levant.12 In Brit-
around the age of 5 years old and again, after the age of 9 years ain, in addition to one outlier individual with ancestry related to

Current Biology 34, 204–212, January 8, 2024 207

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present-day Near Eastern populations in the possible military or slavery. The absence of grave goods and the generally unre-
gladiator cemetery at Driffield Terrace, in present-day York markable nature of his grave prevents assessment of which sce-
(Eboracum, a major urban center and provincial capital),9 nario is most likely. A plausible explanation is that he died while
isotope signatures consistent with continental Europe and the en route somewhere, although this scenario may be weakened
Mediterranean basin have also been reported at other important by the location of his burial one kilometer to the west of a major
urban Roman settlements36,39,40. By contrast, Offord Cluny Roman road connecting Sandy and Godmanchester, Durovigu-
203645 was found in what would have been a rural location, tum (Figure 1A). An alternative hypothesis is that Offord Cluny
albeit within a substantial farmstead that later developed into a 203645 was associated with the farmstead, possibly integrated
villa complex. The skeleton was not recovered from one of the within a rural civilian community.
small formal Romano-British cemeteries found along the modern Whatever the reasons for the journeys Offord Cluny 203645
A14 road, but from an isolated burial that had been placed within took over his short lifetime, his burial highlights the impact that
a former trackway ditch toward the fringes of the farmstead. Iso- the Roman Empire had on rural locations in Britain (and probably
lated burials outside of formal cemeteries in peripheral unfur- elsewhere) in terms of increasing long-distance mobility and
nished graves are a common feature of early-mid Roman farm- introducing genetic ancestry from the far fringes or even regions
steads and villas.41,42 It is usually unclear who was placed in outside of the Roman Empire. Future identification of additional
these isolated burials, though the very act of interment itself individuals with Caucasus- and/or Sarmatian-related ancestry in
does distinguish them, with the majority of the rural population Roman Britain, particularly examples accompanied by grave
during the early-mid Roman period having been subjected goods or from indicative contexts (e.g., military), will offer more
to funerary rites which left little archaeological trace (e.g., insights into how people who carried these ancestries arrived
excarnation). in Britain.
Contributions of Caucasus- or Pontic-Caspian-associated
ancestry, usually admixed with local populations, have been STAR+METHODS
identified in Roman cemeteries in other parts of the empire,
such as in Italy or the Balkans4,5 (Figure 1C). The second century Detailed methods are provided in the online version of this paper
CE witnessed a series of interactions between the Roman Em- and include the following:
pire and the inhabitants of the Caucasus, including a brief period
between 114 and 117 CE when Greater Armenia became a Ro- d KEY RESOURCES TABLE
man province,43 as well as several documented Sarmatian-Alan d RESOURCE AVAILABILITY
incursions into the Roman-controlled South Caucasus.2 In the B Lead contact
northeastern fringes of the empire, the Marcomannic Wars B Materials availability
(166–180 CE) pitted the Romans against Germanic and Sarma- B Data and code availability
tian peoples.1 All of these events could have promoted long-dis- d EXPERIMENTAL MODEL AND STUDY PARTICIPANT DE-
tance mobility of groups or individuals carrying Caucasus- and TAILS
Sarmatian-related ancestry into and within the Roman Empire. B Archaeological context
The age at death (18–25 years old) and history of migration B Skeletal samples
(based both on genetic ancestry and stable isotope evidence) d METHOD DETAILS
we have obtained from Offord Cluny 203645 could be consistent B DNA sampling and sequencing
with this individual having come to Britain as part of a military B Strontium isotopes
movement, either as part of a soldier’s family or as a soldier him- B Oxygen isotopes
self. One possibility, given the radiocarbon date obtained (126– B Carbon and nitrogen isotopes
228 cal. CE; median 176 cal. CE), would be the historically at- d QUANTIFICATION AND STATISTICAL ANALYSIS
tested deployment of Sarmatian cavalry in 175 CE, following B Sequencing data processing and aDNA authentication
Roman emperor Marcus Aurelius’s victory in the Marcomannic B Genotyping and compiled datasets
Wars, as described by the Roman historian Cassius Dio.4,5 In B Population analyses
this scenario, the dietary shifts we see in Offord Cluny 203645
would be explicable if he was associated with groups of Sarma- SUPPLEMENTAL INFORMATION
tians who moved into central Europe before or during the Marco-
Supplemental information can be found online at https://doi.org/10.1016/j.
mannic wars,3 although the plausibility of this interpretation de-
cub.2023.11.049.
pends on whether children were likely to have been part of
movements of Sarmatians across Europe. Little is known about ACKNOWLEDGMENTS
where the 5,500 Sarmatians were stationed in Britain. There are
suggestions of Sarmatian horse equipment from Chesters on We thank National Highways for supporting this study, and Jesse McCabe,
Hadrian’s Wall and epigraphic evidence for them from Ribches- Leo Speidel, and Pooja Swali for helpful discussions. We thank Beata
ter, Bremetennacum Veteranorum in northwest England and Wieczorek-Oleksy from Headland Archaeology for providing the site map
Catterick, Cataractonium in northeast England,4,44 all a consid- and Joe Brock for helping with the graphical abstract. We thank Ron Pinhasi,
Jonathan Pritchard, and co-authors of Antonio et al.12 for making the data
erable distance from the A14 sites in Cambridgeshire.
available ahead of peer-reviewed publication, and the three anonymous re-
Other interpretations that could plausibly account for long-dis- viewers for their constructive comments which helped to improve this paper.
tance movement across the Roman Empire include, although are This work was supported by the European Molecular Biology Organisation,
not limited to, governance of the empire, economic migration, or the Vallee Foundation, the European Research Council (grant no. 852558),

208 Current Biology 34, 204–212, January 8, 2024

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the Wellcome Trust (217223/Z/19/Z), and Francis Crick Institute core funding high mobility. Preprint at bioRxiv. https://doi.org/10.1101/2022.05.15.
(FC001595) from Cancer Research UK, the UK Medical Research Council, 491973.
and the Wellcome Trust. We thank the Advanced Sequencing Facility and Sci- 13. Lazaridis, I., Alpaslan-Roodenberg, S., Acar, A., Açıkkol, A., Agelarakis,
entific Computing at the Francis Crick institute for technical support. For the A., Aghikyan, L., Akyüz, U., Andreeva, D., Andrija
sevic
, G., Antonovic ,
purpose of open access, the author has applied a CC BY public copyright D., et al. (2022). The genetic history of the Southern Arc: A bridge be-
licence to any Author Accepted Manuscript version arising from this tween West Asia and Europe. Science 377, eabm4247.
submission.
14. Lazaridis, I., Nadel, D., Rollefson, G., Merrett, D.C., Rohland, N., Mallick,
S., Fernandes, D., Novak, M., Gamarra, B., Sirak, K., et al. (2016).
AUTHOR CONTRIBUTIONS
Genomic insights into the origin of farming in the ancient Near East.
Nature 536, 419–424.
M.S., T.B., D.W., J. Montgomery, and P.S. designed the study. D.W., J. Mont-
gomery, and P.S. led the research teams. D.W., M.H., A.S., and D.B. identified 15. Gamba, C., Jones, E.R., Teasdale, M.D., McLaughlin, R.L., Gonzalez-
archaeological material and provided interpretation. M.S., T.B., K.A., M.K., Fortes, G., Mattiangeli, V., Domboróczki, L., Ko } vári, I., Pap, I., Anders,
and M.W. extracted and sequenced ancient DNA. M.S., A.G., and C.B. per- A., et al. (2014). Genome flux and stasis in a five millennium transect of
formed bioinformatic processing. M.S. performed statistical genetic analysis. European prehistory. Nat. Commun. 5, 5257.
J. Moore and J. Montgomery conducted stable isotope analyses. M.S., T.B, 16. Yonova-Doing, E., Calabrese, C., Gomez-Duran, A., Schon, K., Wei, W.,
D.W., and P.S. prepared the manuscript. Karthikeyan, S., Chinnery, P.F., and Howson, J.M.M. (2021). An atlas of
mitochondrial DNA genotype-phenotype associations in the UK
DECLARATION OF INTERESTS Biobank. Nat. Genet. 53, 982–993.
17. Olalde, I., Brace, S., Allentoft, M.E., Armit, I., Kristiansen, K., Booth, T.,
The authors declare no competing interests. Rohland, N., Mallick, S., Sze csenyi-Nagy, A., Mittnik, A., et al. (2018).
The Beaker phenomenon and the genomic transformation of northwest
Received: July 19, 2023 Europe. Nature 555, 190–196.
Revised: October 10, 2023
18. Brace, S., Diekmann, Y., Booth, T.J., van Dorp, L., Faltyskova, Z.,
Accepted: November 21, 2023
Rohland, N., Mallick, S., Olalde, I., Ferry, M., Michel, M., et al. (2019).
Published: December 19, 2023
Ancient genomes indicate population replacement in Early Neolithic
Britain. Nat. Ecol. Evol. 3, 765–771.
REFERENCES
19. Sánchez-Quinto, F., Malmström, H., Fraser, M., Girdland-Flink, L.,
1. Fischer, T. (2012). Archaeological evidence of the Marcomannic wars of Svensson, E.M., Simões, L.G., George, R., Hollfelder, N., Burenhult, G.,
Marcus Aurelius (AD 166-80). In A Companion to Marcus Aurelius Noble, G., et al. (2019). Megalithic tombs in western and northern
(Oxford: Wiley-Blackwell), pp. 29–44. Neolithic Europe were linked to a kindred society. Proc. Natl. Acad.
Sci. USA 116, 9469–9474.
2. Gregoratti, L. (2013). The Caucasus: a communication Space between
Nomads and Sedentaries (1st BC-2nd AD). In Le aree montane come 20. Schiffels, S., Haak, W., Paajanen, P., Llamas, B., Popescu, E., Loe, L.,
frontiere. Spazi d’interazione e connettività, S. Magnani, ed. (Roma: Clarke, R., Lyons, A., Mortimer, R., Sayer, D., et al. (2016). Iron Age
Aracne Editrice), pp. 477–493. and Anglo-Saxon genomes from East England reveal British migration
history. Nat. Commun. 7, 10408.
3. Alemany, A. (2000). Sources on the Alans: A Critical Compilation (Leiden:
Brill). 21. Gretzinger, J., Sayer, D., Justeau, P., Altena, E., Pala, M., Dulias, K.,
4. Richmond, I.A. (1945). The Sarmatae, Bremetennacvm Veteranorvm and Edwards, C.J., Jodoin, S., Lacher, L., Sabin, S., et al. (2022). The
the Regio Bremetennacensis. J. Rom. Stud. 35, 15–29. Anglo-Saxon migration and the formation of the early English gene
pool. Nature 610, 112–119.
5. Birley, A.R. (2012). Cassius Dio and the Historia Augusta. In A Companion
to Marcus Aurelius (Wiley-Blackwell), pp. 11–28. 22. Reid, D.J., and Dean, M.C. (2006). Variation in modern human enamel for-
mation times. J. Hum. Evol. 50, 329–346.
6. Skoglund, P., Storå, J., Götherström, A., and Jakobsson, M. (2013).
Accurate sex identification of ancient human remains using DNA shotgun 23. Evans, J.A., Mee, K., Chenery, C.A., Cartwright, C.E., Lee, K.A., and
sequencing. J. Archaeol. Sci. 40, 4477–4482. Marchant, A.P. (2018). User guide for the Biosphere Isotope Domains
GB (Version 1) dataset and web portal. https://nora.nerc.ac.uk/id/
7. Bronk Ramsey, C. (2009). Bayesian Analysis of Radiocarbon Dates.
eprint/520128/1/OR18005.pdf.
Radiocarbon 51, 337–360.
8. Reimer, P.J., Austin, W.E.N., Bard, E., Bayliss, A., Blackwell, P.G., Bronk 24. Evans, J.A., Chenery, C.A., and Montgomery, J. (2012). A summary of
Ramsey, C., Butzin, M., Cheng, H., Edwards, R.L., Friedrich, M., et al. strontium and oxygen isotope variation in archaeological human tooth
(2020). The IntCal20 Northern Hemisphere Radiocarbon Age enamel excavated from Britain. J. Anal. At. Spectrom. 27, 754–764.
Calibration Curve (0–55 cal kBP). Radiocarbon 62, 725–757. 25. Garbaras, A., Skipityte,

_ R., Sapolait
and Remeikis, V.
_ J., E
zerinskis, Z.,
e,
9. Martiniano, R., Caffell, A., Holst, M., Hunter-Mann, K., Montgomery, J., (2019). Seasonal Variation in Stable Isotope Ratios of Cow Milk in Vilnius
Müldner, G., McLaughlin, R.L., Teasdale, M.D., van Rheenen, W., Region, Lithuania. Animals. 9, 69. https://doi.org/10.3390/ani9030069.
Veldink, J.H., et al. (2016). Genomic signals of migration and continuity 26. Crowder, K.D., Montgomery, J., Filipek, K.L., and Evans, J.A. (2020).
in Britain before the Anglo-Saxons. Nat. Commun. 7, 10326. Romans, barbarians and foederati: New biomolecular data and a
10. Damgaard, P.d.B., Marchi, N., Rasmussen, S., Peyrot, M., Renaud, G., possible region of origin for ‘‘Headless Romans’’ and other burials from
Korneliussen, T., Moreno-Mayar, J.V., Pedersen, M.W., Goldberg, A., Britain. J. Archaeol. Sci. Rep. 30, 102180.
Usmanova, E., et al. (2018). 137 ancient human genomes from across 27. Fiorin, E., Moore, J., Montgomery, J., Lippi, M.M., Nowell, G., and Forlin,
the Eurasian steppes. Nature 557, 369–374. P. (2023). Combining dental calculus with isotope analysis in the Alps:
11. Moshkova, M.G. (1995). Late Sarmatian Culture. In Nomads of the New evidence from the Roman and medieval cemeteries of Lamon.
Eurasian Steppes in the Early Iron Age, J. Davis-Kimball, V.A. Bashilov, Quat. Int. 653–654, 89–102.
and L.T. Yablonsky, eds. (Berkeley: Zinat Press), pp. 149–163. 28. Montgomery, J., Evans, J., and Towers, J. (2019). Strontium isotope
12. Antonio, M.L., Weiß, C.L., Gao, Z., Sawyer, S., Oberreiter, V., Moots, analysis. In The Beaker People: Isotopes, mobility and diet, M. Parker
H.M., Spence, J.P., Cheronet, O., Zagorc, B., Praxmarer, E., et al. Pearson, A. Sheridan, M. Jay, A. Chamberlain, M.P. Richards, and J.
(2022). Stable population structure in Europe since the Iron Age, despite Evans, eds. (Oxford: Oxbow), pp. 369–406.

Current Biology 34, 204–212, January 8, 2024 209

 ll
OPEN ACCESS Report
29. Pellegrini, M., Jay, M., and Richards, M.P. (2019). Oxygen isotope anal- A. (2016). Genomic Evidence Establishes Anatolia as the Source of the
ysis. In The Beaker People: Isotopes, mobility and diet, M. Parker European Neolithic Gene Pool. Curr. Biol. 26, 270–275.
Pearson, A. Sheridan, M. Jay, A. Chamberlain, M.P. Richards, and J. 49. Marchi, N., Winkelbach, L., Schulz, I., Brami, M., Hofmanová, Z., Blöcher,
Evans, eds. (Oxbow), pp. 407–424. J., Reyna-Blanco, C.S., Diekmann, Y., Thie ry, A., Kapopoulou, A., et al.
30. Müldner, G. (2013). Stable isotopes and diet: their contribution to (2022). The genomic origins of the world’s first farmers. Cell 185, 1842–
Romano-British research. Antiquity 87, 137–149. 1859.e18.
31. Dean, M.C., and Cole, T.J. (2013). Human life history evolution explains 50. Jones, E.R., Gonzalez-Fortes, G., Connell, S., Siska, V., Eriksson, A.,
dissociation between the timing of tooth eruption and peak rates of Martiniano, R., McLaughlin, R.L., Gallego Llorente, M., Cassidy, L.M.,
root growth. PLoS One 8, e54534. Gamba, C., et al. (2015). Upper Palaeolithic genomes reveal deep roots
32. Dean, C., and Cole, T. (2014). The Timing of our tooth growth is an evolu- of modern Eurasians. Nat. Commun. 6, 8912.
tionary relic. Significance 11, 19–23. 51. Saag, L., Vasilyev, S.V., Varul, L., Kosorukova, N.V., Gerasimov, D.V.,
33. Hillson, S. (1996). Dental Anthropology, Second Edition (Cambridge: Oshibkina, S.V., Griffith, S.J., Solnik, A., Saag, L., D’Atanasio, E., et al.
Cambridge University Press). (2021). Genetic ancestry changes in Stone to Bronze Age transition in
the East European plain. Sci. Adv. 7, eabd6535.
34. Aufderheide, A.C., Rodrı́guez-Martı́n, C., and Langsjoen, O. (2014). The
Cambridge encyclopedia of human paleopathology (Cambridge: 52. Fu, Q., Posth, C., Hajdinjak, M., Petr, M., Mallick, S., Fernandes, D.,
Cambridge University Press). Furtwa€ngler, A., Haak, W., Meyer, M., Mittnik, A., et al. (2016). The ge-
35. Müldner, G., Chenery, C., and Eckardt, H. (2011). The ‘‘Headless netic history of Ice Age Europe. Nature 534, 200–205.
Romans’’: multi-isotope investigations of an unusual burial ground from 53. de Barros Damgaard, P., Martiniano, R., Kamm, J., Vı́ctor Moreno-
Roman Britain. J. Archaeol. Sci. 38, 280–290. Mayar, J., Kroonen, G., Peyrot, M., Barjamovic, G., Rasmussen, S.,
36. Chenery, C., Müldner, G., Evans, J., Eckardt, H., and Lewis, M. (2010). Zacho, C., Baimukhanov, N., et al. (2018). The first horse herders and
Strontium and stable isotope evidence for diet and mobility in Roman the impact of early Bronze Age steppe expansions into Asia. Science
Gloucester, UK. J. Archaeol. Sci. 37, 150–163. 360, eaar7711.

, I., Rohland, N., Mallick, S., Lazaridis, I.,

37. Olalde, I., Carrión, P., Mikic 54. Dulias, K., Foody, M.G.B., Justeau, P., Silva, M., Martiniano, R., Oteo-
Korac , M., Golubovic , S., Petkovic , S., Miladinovic-Radmilovic, N., Garcı́a, G., Fichera, A., Rodrigues, S., Gandini, F., Meynert, A., et al.
et al. (2021). Cosmopolitanism at the Roman Danubian Frontier, Slavic (2022). Ancient DNA at the edge of the world: Continental immigration
Migrations, and the Genomic Formation of Modern Balkan Peoples. and the persistence of Neolithic male lineages in Bronze Age Orkney.
Preprint at bioRxiv. https://doi.org/10.1101/2021.08.30.458211. Proc. Natl. Acad. Sci. USA 119, e2108001119.

38. Antonio, M.L., Gao, Z., Moots, H.M., Lucci, M., Candilio, F., Sawyer, S., 55. González-Fortes, G., Jones, E.R., Lightfoot, E., Bonsall, C., Lazar, C.,
Oberreiter, V., Calderon, D., Devitofranceschi, K., Aikens, R.C., et al. Grandal-d’Anglade, A., Garralda, M.D., Drak, L., Siska, V., Simalcsik,
(2019). Ancient Rome: A genetic crossroads of Europe and the A., et al. (2017). Paleogenomic Evidence for Multi-generational Mixing
Mediterranean. Science 366, 708–714. between Neolithic Farmers and Mesolithic Hunter-Gatherers in the
Lower Danube Basin. Curr. Biol. 27, 1801–1810.e10.
39. Redfern, R.C., Gröcke, D.R., Millard, A.R., Ridgeway, V., Johnson, L.,
and Hefner, J.T. (2016). Going south of the river: A multidisciplinary anal- 56. Krzewin ska, M., Kılınç, G.M., Juras, A., Koptekin, D., Chylen
ski, M.,
ysis of ancestry, mobility and diet in a population from Roman Nikitin, A.G., Shcherbakov, N., Shuteleva, I., Leonova, T., Kraeva, L.,
Southwark, London. J. Archaeol. Sci. 74, 11–22. et al. (2018). Ancient genomes suggest the eastern Pontic-Caspian
steppe as the source of western Iron Age nomads. Sci. Adv. 4, eaat4457.
40. Eckardt, H., Chenery, C., Booth, P., Evans, J.A., Lamb, A., and Müldner,
G. (2009). Oxygen and strontium isotope evidence for mobility in Roman 57. Schlebusch, C.M., Malmström, H., Günther, T., Sjödin, P., Coutinho, A.,
Winchester. J. Archaeol. Sci. 36, 2816–2825. Edlund, H., Munters, A.R., Vicente, M., Steyn, M., Soodyall, H., et al.
41. Pearce, J. (2008). Burial evidence from Roman Britain: the un-numbered (2017). Southern African ancient genomes estimate modern human
dead. In Pour une arche ologie du rite. Nouvelles perspectives de l’arch- divergence to 350,000 to 260,000 years ago. Science 358, 652–655.
ologie fune
e raire, J. Scheid, ed. (Rome: Ecole Francaise de Rome), 58. Lazaridis, I., Patterson, N., Mittnik, A., Renaud, G., Mallick, S., Kirsanow,
pp. 29–42. K., Sudmant, P.H., Schraiber, J.G., Castellano, S., Lipson, M., et al.
42. Smith, A., Allen, M., Brindle, T., Fulford, M., Lodwick, L., and (2014). Ancient human genomes suggest three ancestral populations
Rohnbogner, A. (2018). Life and death in the countryside of Roman for present-day Europeans. Nature 513, 409–413.
Britain (London: Society for the Promotion of Roman Studies). 59. Olalde, I., Allentoft, M.E., Sánchez-Quinto, F., Santpere, G., Chiang,
43. Speidel, M.A. (2021). Provincia Armenia in the Light of the Epigraphic C.W.K., DeGiorgio, M., Prado-Martinez, J., Rodrı́guez, J.A.,
_
Evidence. Electrum. Studia z historii starozytnej 28, 135–150. Rasmussen, S., Quilez, J., et al. (2014). Derived immune and ancestral
pigmentation alleles in a 7,000-year-old Mesolithic European. Nature
44. Eckardt, H. (2014). Objects and Identities in Roman Britain and the North-
507, 225–228.
Western Provinces (Oxford: Oxford University Press).
60. Sikora, M., Pitulko, V.V., Sousa, V.C., Allentoft, M.E., Vinner, L.,
45. Allentoft, M.E., Sikora, M., Sjögren, K.G., Rasmussen, S., Rasmussen,
Rasmussen, S., Margaryan, A., de Barros Damgaard, P., de la Fuente,
M., Stenderup, J., Damgaard, P.B., Schroeder, H., Ahlström, T., Vinner,
C., Renaud, G., et al. (2019). The population history of northeastern
L., et al. (2015). Population genomics of Bronze Age Eurasia. Nature
Siberia since the Pleistocene. Nature 570, 182–188.
522, 167–172.
61. Mallick, S., Micco, A., Mah, M., Ringbauer, H., Lazaridis, I., Olalde, I.,
46. Yaka, R., Mapelli, I., Kaptan, D., Dog u, A., Chylen
ski, M., Erdal, Ö.D.,
Patterson, N., and Reich, D. (2023). The Allen Ancient DNA Resource
Koptekin, D., Vural, K.B., Bayliss, A., Mazzucato, C., et al. (2021).
(AADR): A curated compendium of ancient human genomes. Preprint
Variable kinship patterns in Neolithic Anatolia revealed by ancient ge-
at bioRxiv. https://doi.org/10.1101/2023.04.06.535797.
nomes. Curr. Biol. 31, 2455–2468.e18.
62. The 1000 Genomes Project Consortium (2015). A global reference for hu-
47. Hofmanová, Z., Kreutzer, S., Hellenthal, G., Sell, C., Diekmann, Y., Dı́ez-
man genetic variation. Nature 526, 68–74.
Del-Molino, D., van Dorp, L., López, S., Kousathanas, A., Link, V., et al.
(2016). Early farmers from across Europe directly descended from 63. van Oven, M. (2015). PhyloTree Build 17: Growing the human mitochon-
Neolithic Aegeans. Proc. Natl. Acad. Sci. USA 113, 6886–6891. drial DNA tree. Forensic Sci. Int. Genet. Suppl. Ser. 5, e392–e394.
48. Omrak, A., Günther, T., Valdiosera, C., Svensson, E.M., Malmström, H., 64. Gansauge, M.T., Aximu-Petri, A., Nagel, S., and Meyer, M. (2020).
Kiesewetter, H., Aylward, W., Storå, J., Jakobsson, M., and Götherström, Manual and automated preparation of single-stranded DNA libraries for

210 Current Biology 34, 204–212, January 8, 2024

 ll
Report OPEN ACCESS

the sequencing of DNA from ancient biological remains and other sour- 85. Kircher, M., Sawyer, S., and Meyer, M. (2012). Double indexing over-
ces of highly degraded DNA. Nat. Protoc. 15, 2279–2300. comes inaccuracies in multiplex sequencing on the Illumina platform.
65. Gansauge, M.T., and Meyer, M. (2013). Single-stranded DNA library Nucleic Acids Res. 40, e3.
preparation for the sequencing of ancient or damaged DNA. Nat. 86. Font, L., Davidson, J.P., Pearson, D.G., Nowell, G.M., Jerram, D.A., and
Protoc. 8, 737–748. Ottley, C.J. (2008). Sr and Pb Isotope Micro-analysis of Plagioclase
66. Fellows Yates, J.A., Lamnidis, T.C., Borry, M., Andrades Valtueña, A., Crystals from Skye Lavas: an Insight into Open-system Processes in a
Fagerna €s, Z., Clayton, S., Garcia, M.U., Neukamm, J., and Peltzer, A. Flood Basalt Province. J. Petrol. 49, 1449–1471.
(2021). Reproducible, portable, and efficient ancient genome reconstruc- 87. Coplen, T.B., Kendall, C., and Hopple, J. (1983). Comparison of stable
tion with nf-core/eager. PeerJ 9, e10947. isotope reference samples. Nature 302, 236–238.
67. Chen, S., Zhou, Y., Chen, Y., and Gu, J. (2018). fastp: an ultra-fast all-in- 88. Beaumont, J., Gledhill, A., and Montgomery, J. (2014). Isotope analysis of
one FASTQ preprocessor. Bioinformatics 34, i884–i890. incremental human dentine: towards higher temporal resolution. Bull. Int.
68. Schubert, M., Lindgreen, S., and Orlando, L. (2016). AdapterRemoval v2: Assoc. Paleodontol. 8, 212–223.
rapid adapter trimming, identification, and read merging. BMC Res. 89. Mathieson, I., Lazaridis, I., Rohland, N., Mallick, S., Patterson, N.,
Notes 9, 88. Roodenberg, S.A., Harney, E., Stewardson, K., Fernandes, D., Novak,
69. Li, H., and Durbin, R. (2009). Fast and accurate short read alignment with M., et al. (2015). Genome-wide patterns of selection in 230 ancient
Burrows-Wheeler transform. Bioinformatics 25, 1754–1760. Eurasians. Nature 528, 499–503.
€ger, G., Herbig, A., Seitz, A., Kniep, C., Krause, J., and
70. Peltzer, A., Ja 90. Patterson, N., Isakov, M., Booth, T., Büster, L., Fischer, C.-E., Olalde, I.,
Nieselt, K. (2016). EAGER: efficient ancient genome reconstruction. Ringbauer, H., Akbari, A., Cheronet, O., Bleasdale, M., et al. (2022).
Genome Biol. 17, 60. Large-scale migration into Britain during the Middle to Late Bronze
Age. Nature 601, 588–594.
71. Korneliussen, T.S., Albrechtsen, A., and Nielsen, R. (2014). ANGSD:
Analysis of Next Generation Sequencing Data. BMC Bioinf. 15, 356. 91. Marcus, J.H., Posth, C., Ringbauer, H., Lai, L., Skeates, R., Sidore, C.,
€ngler, A., Olivieri, A., Chiang, C.W.K., et al. (2020).
Beckett, J., Furtwa
72. Renaud, G., Slon, V., Duggan, A.T., and Kelso, J. (2015). Schmutzi: esti-
Genetic history from the Middle Neolithic to present on the
mation of contamination and endogenous mitochondrial consensus call-
Mediterranean island of Sardinia. Nat. Commun. 11, 939.
ing for ancient DNA. Genome Biol. 16, 224.
92. De Angelis, F., Romboni, M., Veltre, V., Catalano, P., Martı́nez-Labarga,
73. Neukamm, J., Peltzer, A., and Nieselt, K. (2021). DamageProfiler: fast
C., Gazzaniga, V., and Rickards, O. (2022). First Glimpse into the
damage pattern calculation for ancient DNA. Bioinformatics 37,
Genomic Characterization of People from the Imperial Roman
3652–3653.
Community of Casal Bertone (Rome, First-Third Centuries AD). Genes
74. Ralf, A., Montiel González, D., Zhong, K., and Kayser, M. (2018). Yleaf:
13, 136.
Software for Human Y-Chromosomal Haplogroup Inference from Next-
93. Fernandes, D.M., Mittnik, A., Olalde, I., Lazaridis, I., Cheronet, O.,
Generation Sequencing Data. Mol. Biol. Evol. 35, 1291–1294.
Rohland, N., Mallick, S., Bernardos, R., Broomandkhoshbacht, N.,
75. Li, H., Handsaker, B., Wysoker, A., Fennell, T., Ruan, J., Homer, N.,
Carlsson, J., et al. (2020). The spread of steppe and Iranian-related
Marth, G., Abecasis, G., and Durbin, R.; 1000 Genome Project Data
ancestry in the islands of the western Mediterranean. Nat. Ecol. Evol.
Processing Subgroup (2009). The Sequence Alignment/Map format
4, 334–345.
and SAMtools. Bioinformatics 25, 2078–2079.
94. Haber, M., Doumet-Serhal, C., Scheib, C.L., Xue, Y., Mikulski, R.,
76. Weissensteiner, H., Pacher, D., Kloss-Brandsta €tter, A., Forer, L., Specht,
Martiniano, R., Fischer-Genz, B., Schutkowski, H., Kivisild, T., and
G., Bandelt, H.-J., Kronenberg, F., Salas, A., and Schönherr, S. (2016).
Tyler-Smith, C. (2019). A Transient Pulse of Genetic Admixture from the
HaploGrep 2: mitochondrial haplogroup classification in the era of
Crusaders in the Near East Identified from Ancient Genome
high-throughput sequencing. Nucleic Acids Res. 44, W58–W63.
Sequences. Am. J. Hum. Genet. 104, 977–984.
77. Purcell, S., Neale, B., Todd-Brown, K., Thomas, L., Ferreira, M.A.R.,
95. Haber, M., Nassar, J., Almarri, M.A., Saupe, T., Saag, L., Griffith, S.J.,
Bender, D., Maller, J., Sklar, P., de Bakker, P.I.W., Daly, M.J., and
Doumet-Serhal, C., Chanteau, J., Saghieh-Beydoun, M., Xue, Y., et al.
Sham, P.C. (2007). PLINK: a tool set for whole-genome association
(2020). A Genetic History of the Near East from an aDNA Time Course
and population-based linkage analyses. Am. J. Hum. Genet. 81,
Sampling Eight Points in the Past 4,000 Years. Am. J. Hum. Genet.
559–575.
107, 149–157.
78. Patterson, N., Price, A.L., and Reich, D. (2006). Population structure and €rve, M., Saag, L., Scheib, C.L., Pathak, A.K., Montinaro, F., Pagani, L.,
96. Ja
eigenanalysis. PLoS Genet. 2, e190.
Flores, R., Guellil, M., Saag, L., Tambets, K., et al. (2019). Shifts in the
79. Patterson, N., Moorjani, P., Luo, Y., Mallick, S., Rohland, N., Zhan, Y., Genetic Landscape of the Western Eurasian Steppe Associated with
Genschoreck, T., Webster, T., and Reich, D. (2012). Ancient admixture the Beginning and End of the Scythian Dominance. Curr. Biol. 29,
in human history. Genetics 192, 1065–1093. 2430–2441.e10.
80. Skoglund, P., Mallick, S., Bortolini, M.C., Chennagiri, N., Hünemeier, T., 97. Narasimhan, V.M., Patterson, N.J., Moorjani, P., Lazaridis, I., Lipson, M.,
Petzl-Erler, M.L., Salzano, F.M., Patterson, N., and Reich, D. (2015). Mallick, S., Rohland, N., Bernardos, R., Kim, A.M., Nakatsuka, N., et al.
Genetic evidence for two founding populations of the Americas. Nature (2018). The Genomic Formation of South and Central Asia. Preprint at
525, 104–108. bioRxiv. https://doi.org/10.1101/292581.
81. Gustafson, G., and Koch, G. (1974). Age estimation up to 16 years of age 98. Olalde, I., Mallick, S., Patterson, N., Rohland, N., Villalba-Mouco, V.,
based on dental development. Odontol. Revy 25, 297–306. Silva, M., Dulias, K., Edwards, C.J., Gandini, F., Pala, M., et al. (2019).
82. Anderson, D.L., Thompson, G.W., and Popovich, F. (1976). Age of attain- The genomic history of the Iberian Peninsula over the past 8000 years.
ment of mineralization stages of the permanent dentition. J. Forensic Sci. Science 363, 1230–1234.
21, 191–200. 99. Posth, C., Zaro, V., Spyrou, M.A., Vai, S., Gnecchi-Ruscone, G.A., Modi,
83. Pinhasi, R., Fernandes, D., Sirak, K., Novak, M., Connell, S., Alpaslan- €gele, K., Vågene, Å.J., et al. (2021). The
A., Peltzer, A., Mötsch, A., Na
Roodenberg, S., Gerritsen, F., Moiseyev, V., Gromov, A., Raczky, P., origin and legacy of the Etruscans through a 2000-year archeogenomic
et al. (2015). Optimal Ancient DNA Yields from the Inner Ear Part of the time transect. Sci. Adv. 7, eabi7673.
Human Petrous Bone. PLoS One 10, e0129102. 100. Scorrano, G., Viva, S., Pinotti, T., Fabbri, P.F., Rickards, O., and
84. Rohland, N., Glocke, I., Aximu-Petri, A., and Meyer, M. (2018). Extraction Macciardi, F. (2022). Bioarchaeological and palaeogenomic portrait of
of highly degraded DNA from ancient bones, teeth and sediments for two Pompeians that died during the eruption of Vesuvius in 79 AD. Sci.
high-throughput sequencing. Nat. Protoc. 13, 2447–2461. Rep. 12, 6468.

Current Biology 34, 204–212, January 8, 2024 211

 ll
OPEN ACCESS Report
€nder, M., Palstra, F., Lazaridis, I., Pilipenko, A., Hofmanová, Z.,
101. Unterla (2022). Ancient genomes reveal origin and rapid trans-Eurasian migration
Groß, M., Sell, C., Blöcher, J., Kirsanow, K., Rohland, N., et al. (2017). of 7th century Avar elites. Cell 185, 1402–1413.e21.
Ancestry and demography and descendants of Iron Age nomads of the
Eurasian Steppe. Nat. Commun. 8, 14615.
ic
104. Freilich, S., Ringbauer, H., Los, D., Novak, M., Pavic , D.T., Schiffels, S.,
102. Veeramah, K.R., Rott, A., Groß, M., van Dorp, L., López, S., Kirsanow, K., and Pinhasi, R. (2021). Reconstructing genetic histories and social orga-
Sell, C., Blöcher, J., Wegmann, D., Link, V., et al. (2018). Population nisation in Neolithic and Bronze Age Croatia. Sci. Rep. 11, 16729.
genomic analysis of elongated skulls reveals extensive female-biased
immigration in Early Medieval Bavaria. Proc. Natl. Acad. Sci. USA 115, 105. Skoglund, P., Thompson, J.C., Prendergast, M.E., Mittnik, A., Sirak, K.,
3494–3499. Hajdinjak, M., Salie, T., Rohland, N., Mallick, S., Peltzer, A., et al.
103. Gnecchi-Ruscone, G.A., Sze csenyi-Nagy, A., Koncz, I., Csiky, G., Rácz, (2017). Reconstructing Prehistoric African Population Structure. Cell
Z., Rohrlach, A.B., Brandt, G., Rohland, N., Csáky, V., Cheronet, O., et al. 171, 59–71.e21.

212 Current Biology 34, 204–212, January 8, 2024

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STAR+METHODS

KEY RESOURCES TABLE

REAGENT or RESOURCE SOURCE IDENTIFIER

Biological samples
Archeological samples: right temporal bone; This study Skeleton (Sk) 203645 (Additional identifiers:
second right mandibular molar; Burial 20.507; C10271)
second right maxillary molar
Chemicals, peptides, and recombinant proteins
T4 DNA Ligase (5 U/mL) Fisher Scientific Cat# EL0012
FastAP Thermosensitive Alkaline Fisher Scientific Cat# EF0651
Phosphatase (1 U/mL)
Klenow Fragment (10U/ul) Fisher Scientific Cat# EP0052
T4 Polynucleotide Kinase (10 U/mL) Fisher Scientific Cat# EK0031
T4 RNA Ligase Reaction Buffer NEB Cat# B0216
ATP Solution (100 mM) Fisher Scientific Cat# R0441
dNTP Mix (25 mM each) VWR Cat# 733-1854
Dynabeads MyOne Streptavidin C1 beads Thermo Fisher Scientific Cat# 65002
G-Biosciences Silica Magnetic Beads VWR Cat# 786-915
AccuPrime Pfx DNA Polymerase Thermo Fisher Scientific Cat# 12344024
Sera-Mag SpeedBeads, magnetic Sigma-Aldrich Cat# GE65152105050250
carboxylate-modified microparticles
Herculase II Fusion DNA Polymerase Agilent Cat# 600679
pUC19 vector NEB Cat# N3041S
Hydrochloric acid, >37% (0.5 M Solution) Sigma-Aldrich Cat# 30721-M
6 M Hydrochloric acid Romil Distilled and titrated in-house
3 M Nitric acid Romil Distilled and titrated in-house
Sr-Spec Resin Triskem Cat# SR-B25-S
Critical commercial assays
MinElute PCR Purification Kit Qiagen Cat# 28004
High Pure Viral Nucleic Acid Roche Cat# 05114403001
Large Volume Kit
Maxima Probe qPCR Master Mix Fisher Scientific Cat# K0262
Agilent DNA 1000 Kit Agilent Cat# 5067-1504
Deposited data
Offord Cluny Sk203645 (Burial 20.507, C10271): This study https://www.ebi.ac.uk/ena/browser/view/PRJEB67353
FASTQ files and mapped BAM file
Human reference genome Genome Reference https://www.ncbi.nlm.nih.gov/grc/human
NCBI build 37, GRCh37 Consortium
Comparison shotgun data Allentoft et al.45 https://www.ebi.ac.uk/ena/browser/view/PRJEB9021
Comparison shotgun data Yaka et al.46 https://www.ebi.ac.uk/ena/browser/view/PRJEB39316
Comparison shotgun data Hofmanová et al.47 https://www.ebi.ac.uk/ena/browser/view/PRJEB11848
Comparison shotgun data Omrak et al.48 https://www.ebi.ac.uk/ena/browser/view/PRJEB12155
Comparison shotgun data Antonio et al.12 https://www.ebi.ac.uk/ena/browser/view/PRJEB53564
Comparison shotgun data Gamba et al.15 https://www.ebi.ac.uk/ena/browser/view/PRJNA240906
Comparison shotgun data Marchi et al.49 https://www.ebi.ac.uk/ena/browser/view/PRJEB50857
Comparison shotgun data Jones et al.50 https://www.ebi.ac.uk/ena/browser/view/PRJEB11364
Comparison shotgun data Saag et al.51 https://www.ebi.ac.uk/ena/browser/view/PRJEB40698
Comparison shotgun data Fu et al.52 https://www.ebi.ac.uk/ena/browser/view/PRJEB13123
Comparison shotgun data de Barros Damgaard et al.53 https://www.ebi.ac.uk/ena/browser/view/PRJEB26349
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Continued
REAGENT or RESOURCE SOURCE IDENTIFIER
20
Comparison shotgun data Schiffels et al. https://www.ebi.ac.uk/ena/browser/view/PRJEB6915
Comparison shotgun data Martiniano et al.9 https://www.ebi.ac.uk/ena/browser/view/PRJEB11004
Comparison shotgun data Dulias et al.54 https://www.ebi.ac.uk/ena/browser/view/PRJEB46830
Comparison shotgun data González-Fortes et al.55 https://www.ebi.ac.uk/ena/browser/view/PRJEB20616
Comparison shotgun data Antonio et al.38 https://www.ebi.ac.uk/ena/browser/view/PRJEB32566
Comparison shotgun data de Barros Damgaard et al.10 https://www.ebi.ac.uk/ena/browser/view/PRJEB20658
Comparison shotgun data ska et al.56
Krzewin https://www.ebi.ac.uk/ena/browser/view/PRJEB27628
Comparison shotgun data Schlebusch et al.57 https://www.ebi.ac.uk/ena/browser/view/PRJEB22660
Comparison shotgun data Lazaridis et al.58 https://www.ebi.ac.uk/ena/browser/view/PRJEB6272
Comparison shotgun data Olalde et al.59 https://www.ebi.ac.uk/ena/browser/view/PRJNA230689
Comparison shotgun data Brace et al.18 https://www.ebi.ac.uk/ena/browser/view/PRJEB31249
Comparison shotgun data Sikora et al.60 https://www.ebi.ac.uk/ena/browser/view/PRJEB29700
‘‘Allen Ancient DNA Resource’’ v.54 Mallick et al.61 https://dataverse.harvard.edu/dataset.xhtml?
persistentId=doi:10.7910/DVN/FFIDCW
1000 Genomes Project (1KGP) phase 3 The 1000 Genomes https://www.internationalgenome.org/category/phase-3/
Project Consortium62
YFull YTree v.11.01.00 N/A https://www.yfull.com/tree/
ISOGG Y-DNA Haplogroup Tree 2019–2020 N/A https://isogg.org/tree/
PhyloTree v.17 van Oven63 https://www.phylotree.org/index.htm
Oligonucleotides
ssDNA library preparation oligonucleotides Gansauge et al.64; N/A
Sigma-Aldrich
CL304, positive control template Gansauge et al.64; N/A
Sigma-Aldrich
P5 and P7 index primers Gansauge and Meyer65; N/A
Sigma-Aldrich
IS5/IS5 biotinylated and IS6, forward Gansauge et al.64; N/A
and reverse primers Sigma-Aldrich
qPCR standard, forward and reverse Gansauge et al.64; N/A
primers and qPCR probes Sigma-Aldrich
forward and reverse primers for Gansauge et al.64; N/A
preparing gel markers Sigma-Aldrich
CL72, sequencing read 1 primer Gansauge et al.64; N/A
for ssDNA libraries Sigma-Aldrich
Software and algorithms
nf-core/eager v.2.3.3 Fellows Yates et al.66 https://nf-co.re/eager/2.3.3
fastp v.0.20.1 Chen et al.67 https://github.com/OpenGene/fastp
AdapterRemoval v2.3.1 Schubert et al.68 https://github.com/MikkelSchubert/adapterremoval
bwa v.0.7.17-r1188 Li and Durbin69 https://github.com/lh3/bwa/releases/tag/v0.7.17
Dedup v.0.12.8 Peltzer et al.70 https://github.com/apeltzer/DeDup/releases/tag/0.12.8
ry_compute.py Skoglund et al.6 https://github.com/pontussk/ry_compute
ANGSD v.0.933 Korneliussen et al.71 http://www.popgen.dk/angsd/index.php/ANGSD
schmutzi v.1.5.6 Renaud et al.72 https://github.com/grenaud/schmutzi
DamageProfiler v.1.1 Neukamm et al.73 https://github.com/Integrative-Transcriptomics/
DamageProfiler
Yleaf v.3.1 Ralf et al.74 https://github.com/genid/Yleaf
samtools v.1.3.1 Li et al.75 https://www.htslib.org/download/
Haplogrep2 Weissensteiner et al.76 https://haplogrep.i-med.ac.at/haplogrep2
sequenceTools v.1.5.2 N/A https://github.com/stschiff/sequenceTools
PLINK v.1.9 Purcell et al.77 https://www.cog-genomics.org/plink/
EIGENSOFT v.6.1.4 Patterson et al.78 https://github.com/DReichLab/EIG
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REAGENT or RESOURCE SOURCE IDENTIFIER
79
ADMIXTOOLS v.5.0 Patterson et al. https://github.com/DReichLab/AdmixTools
qpAdm_wrapper.py N/A https://github.com/pontussk/qpAdm_wrapper
POPSTATS Skoglund et al.80 https://github.com/pontussk/popstats

RESOURCE AVAILABILITY

Lead contact
Further information and requests for resources should be directed to and will be fulfilled by the lead contact, Pontus Skoglund
(pontus.skoglund@crick.ac.uk).

Materials availability
This study did not generate new unique reagents.

Data and code availability

d Sequencing data (FASTQ and BAM files) are available on ENA: PRJEB67353.
d This paper does not report original code.
d Any additional information required to reanalyze the data reported in this paper is available from the lead contact upon request.

EXPERIMENTAL MODEL AND STUDY PARTICIPANT DETAILS

Archaeological context
Between 2016 and 2019 MOLA Headland Infrastructure excavated a series of multiperiod sites in Cambridgeshire, eastern England
on behalf of National Highways as part of the A14 Cambridge-Huntingdon improvement scheme. Amongst other features, these ex-
cavations provided evidence of a well-populated rural Roman landscape comprising a series of complex farmsteads, associated
small cemeteries, villa sites, extensive field systems and isolated human burials. Here, we present genetic and isotopic evidence
of an outlier individual whose remains were recovered from a farmstead (Settlement 2 within the River Great Ouse Landscape Block
of excavations) on the floodplain and gravel terrace of the River Great Ouse, north of the village of Offord Cluny during the A14 ex-
cavations (Figure 1A).
The skeletal remains were recovered from an isolated inhumation. The body appeared to have been laid carefully, slightly flexed on
its left side in a north-south orientation with the head to the south and with the hands crossed in front of the upper legs (Figure S1A).
While there was no evidence of a wrapping or shroud, there may have been some constriction of the body, particularly at the hands
and the knees. The proximity of the hands suggests they may have been deliberately placed, but it is not possible to say whether they
were wrapped or bound. Post-depositional movement of the upper limbs, probably caused by slumping within the grave, has caused
some loss of articulation in the area of the right wrist which may both reflect and mask the original position of the right wrist and hand.
There was no detectable grave cut and no grave goods, although any perishable items would not have survived.

Skeletal samples
Skeleton 203645 (Burial 20.507; Crick ancient genomics lab ID: C10271) comprised the remains of a young adult (aged 18–25 years).
Age estimation was based on observations of dental development and epiphyseal fusion.81,82 The bone was moderately-well pre-
served but the spine, pelvis and lower limbs were degraded and fragmented, which prevented estimation of sex from dimorphic fea-
tures of the skull and pelvis. Linear enamel hypoplastic defects were observed in nine teeth, probably occurring around the age of
5
years based on their location.
We collected the right petrous temporal bone from this individual for aDNA analysis, and the second right mandibular molar tooth
for stable isotope analysis. In addition, the second right maxillary molar was radiocarbon dated to 1867 ± 16 BP (SUERC-105720
(GU61561)) at the Scottish Universities Environmental Research Centre AMS Laboratory, corresponding to 126–228 cal. CE
(95.4% probability) after calibration with OxCal v4.47 using IntCal208 (Figure 1B). Minimally-destructive sampling for aDNA analysis
followed guidelines issued by the Department for Culture, Media and Sport (DCMS) and the Advisory Panel on the Archaeology of
Burials in England (APABE) (apabe.archaeologyuk.org).

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METHOD DETAILS

DNA sampling and sequencing

DNA sampling and pre-amplification protocols were performed in specialized clean rooms at the Francis Crick Institute. We drilled
multiple subsamples of fine bone powder from the cochlear portion of the petrous bone83 using a Emax EVOlution (EV410) micro-
motor system with disposable carbide round burs.
We extracted DNA from a subsample of 18.60 mg of bone powder (using 700 mL of lysis buffer),84 and prepared double-indexed
single-stranded (ss) DNA libraries64,85 without performing any UDG-treatment, using automated liquid-handling systems (Agilent
Bravo Workstations). We included negative extraction and library controls to rule out contamination arising during lab procedures.
Libraries (including negative controls) were initially screened in an Illumina HiSeq 4000 instrument, resulting in
2.6M paired-end (PE)
reads of 100 bp. Following assessment of DNA preservation, we re-sequenced the library twice on the Illumina NovaSeq S4 platform,
for a total of
1.9 billion PE reads, using PE sequencing for 100 cycles (for one of the sequencing rounds we subjected the library to a
gel-excision protocol64 to remove DNA sequences <35 bp and >150 bp).

Strontium isotopes
Core enamel samples (
5 mg) were prepared for strontium (Sr) isotope analysis using column chemistry methods86 at the Arthur
Holmes Isotope Geology Laboratory (AHIGL), Durham University. Samples were digested overnight in 3M HNO3 on a hotplate at
100 C before being loaded onto cleaned and preconditioned columns containing Eichrom strontium-specific resin. A purified Sr frac-
tion was eluted from the column in 400 mL H2O and acidified with 15.5M HNO3 to yield a 3% HNO3 solution. Samples were aspirated
using an ESI PFA-50 nebulizer coupled to a Glass Expansion Cinnabar micro-cyclonic spraychamber. Sr isotopes were measured
using a static multi-collection routine with each measurement comprising a single block of 50 cycles with and integration time of 4s
per cycle (total analysis time
3.5 mins). Instrumental mass bias was corrected for using an 88Sr/86Sr ratio of 8.375209 (the reciprocal
of the more commonly used 86Sr/88Sr ratio of 0.1194) and an exponential law. Corrections for isobaric interferences from Rb and Kr
on 87Sr and 86Sr were performed using 85Rb and 83Kr as the monitor masses but were insignificant. In all samples the 85Rb intensity
was < 1mV with an 85Rb/88Sr ratio of < 0.0003 (average 0.0001). 83Kr was between 0.32 and 0.39mV in all samples. Samples were
measured during a single analytical session during which the average 87Sr/86Sr ratio and reproducibility for the international isotope
reference material NBS987 was 0.710269 ± 0.000013 (2s; n = 12). Maximum error based on internal precision of individual analysis
and analytical reproducibility of the reference material is 0.000013 (2s). Sr isotope data for samples is normalized to an ‘accepted’
value for NBS987 of 0.71024.

Oxygen isotopes
Core enamel samples (
15 mg) were transferred to Iso Analytical for stable isotope analysis where samples were weighed into Ex-
etainer tubes and flushed with 99.995% helium. Carbonate in the samples was converted to CO2 by adding phosphoric acid and
letting the samples sit overnight for the reaction to occur. Reference materials (IA-R022, NBS-18, and IA-R066) were prepared along
the same methods. CO2 from the samples was then analyzed by Continuous Flow-Isotope Ratio Mass Spectrometry (CF-IRMS). The
CO2 was sampled from the Exetainer tubes into a continuously flowing He stream using a double holed needle. The CO2 was resolved
on a packed column gas chromatograph and the resultant chromatographic peak carried forward into the ion source of a Europa
Scientific 20-20 IRMS where it was ionized and accelerated. Gas species of different mass were separated in a magnetic field
then simultaneously measured using a Faraday cup collector array to measure the isotopomers of CO2 at m/z 44, 45, and 46. The
phosphoric acid used for digestion was prepared in accordance with Coplen et al. (1983)87 and was injected through the septum
into the vials. 20% of samples were run in duplicate.

Carbon and nitrogen isotopes

A dentine sample was collected from the root of a second molar and collagen extracted for incremental carbon and nitrogen isotope
analysis following the Beaumont et al. (2014)88 method. Each increment within the dietary profile constitutes a running average (rather
than a discrete snapshot of diet) due to the orientation of the dentine incremental layers and how many are included in each incre-
ment. In human molar teeth, the orientation of these layers is relatively horizontal in the tooth crown (increments 1–6) and becomes
more vertical in the tooth root (increments 7–15), suggesting temporal resolution may be higher in the crown than in the root. For the
second molar the peak velocity is likely to be during increments 1–2 within the crown and increments 7–9 within the root.31,32
Extracted collagen was weighed into tin capsules and measured in duplicate using a Thermo Scientific Delta V Advantage isotope
ratio mass spectrometer in the Stable Isotope Biogeochemistry Laboratory (SIBL), Durham University. Calibration using internal
reference samples (e.g., Glutamic Acid, Glycine, SPAR and Urea) and international reference standards (e.g., USGS 24, USGS
40, IAEA 600, IAEA N1, IAEA N2) determined a standard deviation of ±0.1& (1s) for collagen carbon and nitrogen isotopes. Replicate
analysis of collagen samples averaged a standard deviation of ±0.2& (1s).

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QUANTIFICATION AND STATISTICAL ANALYSIS

Sequencing data processing and aDNA authentication

Sequencing data were processed using nf-core/eager66 v.2.3.3. We preprocessed PE sequencing reads with fastp67 v.0.20.1, fol-
lowed by PE merging and filtering for minimum read length of 35 bp with AdapterRemoval68 v2.3.1 (--collapse, --preserve5p,
--trimns, --trimqualities, --minlength 35, --minquality 20, --minadapteroverlap 1). For libraries sequenced on the Illumina NovaSeq
S4 platform, we performed lane merging before mapping to the human reference genome (hs37d5) using bwa69 v.0.7.17-r1188
aln (-n 0.01, -l 1024, -k 2) and samse. We removed PCR duplicates with Dedup70 v.0.12.8. To generate files containing only mitochon-
drial DNA (mtDNA) reads, we realigned mapped human reads to rCRS (GenBank: NC_012920).
We assigned the libraries as karyotypically male (XY).6 We estimated contamination on the X-chromosome using ANGSD71
v.0.933, and assessed mtDNA contamination using schmutzi72 v.1.5.6 (contDeam.pl --library single) (Data S1A). We merged BAM
files using samtools75 v.1.3.1 merge, and subsequently removed duplicates (Dedup -m), resulting in a final average nuclear coverage
of
5.4x (
4.12x after filtering for mapping quality (MQ) > 30). We used DamageProfiler73 v.1.1 (-sslib) to assess 50 - and 30 -end C>T
substitutions (Figure S1B).
We classified the Y-chromosome lineage using Yleaf74 v.3.1 (-r3, -q30, -dh, -hc) and cross-checked against YFull YTree v.11.01.00
(https://www.yfull.com/tree/) and ISOGG Y-DNA Haplogroup Tree 2019–2020 (https://isogg.org/tree/). For mtDNA haplogroup clas-
sification we used Haplogrep276 based on PhyloTree63 v.17, restricting the data to sites covered by at least four sequencing reads
with MQ > 30 and base quality >30, and allele frequency >0.90.

Genotyping and compiled datasets

We used samtools mpileup (-R, -B, -q30, -Q30) and pileupCaller with the options --randomHaploid and --singleStrandMode (sequen-
ceTools v.1.5.2; https://github.com/stschiff/sequenceTools) to call pseudo-haploid autosomal SNPs overlapping with with the
‘1240k’ panel89 and with
3,868,200 biallelic transversions with 1% minor allele frequency (maf) on the 1000 Genomes Project
(1KGP) phase 3 global panel,62 hereafter referred to as ‘1KGP transversion sites’ (SNP list was generated using PLINK v.1.977
--biallelic-only strict, --maf 0.01).
We extracted genotypes reported in the ‘Allen Ancient DNA Resource’61 v.54 (https://doi.org/10.7910/DVN/FFIDCW). We selected
individuals from England dating to the Iron Age and Roman period, individuals with latitude values between 30 and 64, longitude be-
tween 20 and 60 and mean date between 2000–1475 BP (as reported in the ‘Allen Ancient DNA Resource’ v.54 dataset), but
excluding individuals from early mediaeval contexts. Following a preliminary PCA analysis, we also selected individuals associated
with Sarmatian contexts and individuals from the Caucasus dating to the Late Bronze Age and Iron Age. We retained only unrelated
individuals with >35,000 SNPs overlapping with the ‘1240k’ panel and >20,000 SNPs overlapping with the Affymetrix Human Origins
(HO) array and with no evidence of contamination. We removed close relatives by keeping the individual with the highest number of
genotyped SNPs. The final ‘1240k dataset’ comprised 677 previously reported individuals.9,10,12,13,20,38,45,54,56,89,90–104
We compiled an additional dataset comprising 128 published individuals with whole-genome shotgun data available that we gen-
otyped with samtools mpileup and pileupCaller --randomHaploid using the ‘1KGP transversion sites’ list, as described above (Data
S2D). This comprised a subset of the individuals included in the ‘1240k dataset’ plus additional outgroup and reference popula-
tions15,45,46–53,55,57–60 and was used for all population analyses except PCA.

Population analyses
We used smartpca with options shrinkmode: YES and lsqproject: YES (EIGENSOFT78 v.6.1.4) to project Offord Cluny 203645 along-
side 677 previously published ancient individuals (‘1240k dataset’) on Principal Components (PCs) computed using
600k SNPs
from the HO array genotyped in 1388 present-day individuals from Europe, the Near East and the Caucasus14,58,79 (Data S2E).
We first ran qpAdm framework using a wrapper based on ADMIXTOOLS79 v.5.0 (https://github.com/pontussk/qpAdm_wrapper),
adapting a model optimized for post-Bronze Age Britain,90 with a fixed set of outgroups (ancient sub-Saharan African individuals
(South_Africa_400BP, n = 4), individuals genetically similar to Iron Gates Mesolithic Hunter-gatherers (n = 3), Anatolia Neolithic in-
dividuals (Anatolia_N, n = 18), and Afanasievo individuals (n = 4)) and three distal sources: Western European Hunter-Gatherers
(WHG, n = 7), Neolithic individuals from southeast Europe (Balkan_N, representing European Early Farmers (EEFs) ancestry,
n = 9) and Yamnaya individuals (representing Steppe-associated ancestry, n = 7) (Data S2F). This analysis showed that Offord Cluny
203645 did not harbor WHG-related ancestry (p = 1.65E10) that is otherwise present in the majority of sampled individuals from
post-Bronze Age Western and Central Europe,17,90 and observed in proportions ranging from 15.0 to 21.5% in all non-outlier indi-
viduals from the Driffield Terrace cemetery (Figure S2B and Data S2F). Following this result, we then tested other distal 2-source
models (--sources 2), using a rotating approach105 through a list of reference populations comprising the outgroups and sources
in the previous model plus Caucasus Hunter-Gatherers (CHG, n = 2) and Eastern European Hunter-gatherers (EHG, n = 3) (Figure S2C
and Data S2G).
To find more proximal sources of ancestry, we tested different qpWave (--qpwave –sources 1) and qpAdm (--sources 2) models
using a rotating approach on a selection of West Eurasian populations and additional outgroups (for a total of 4 different reference
lists): South_Africa_400BP (n = 4), Yana_UP (n = 2), Lithuania_Marvele (n = 4), Portugal_LateRoman (n = 5), Italy_ImperialRoman
(n = 20), England_IA (n = 5) or England_Roman (n = 6), Russia_Sarmatian_PonticSteppe (n = 7), Russia_Sarmatian_SouthernUrals
(n = 4), Russia_Sarmatian_Alan (n = 5), Armenia_LBA (n = 7), Armenia_Antiquity (n = 6). We confirmed that none of the Sarmatian

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groups formed a clade with each other (Data S2B). Armenia_LBA was excluded when testing more temporally proximal models. All
tested models with different reference lists are shown in Data S2B and S2C.
We ran f4-statistics using POPSTATS80 (--f4, --haploidize, --informative) to untangle patterns of shared genetic drift amongst
ancient individuals from Roman Britain (Offord Cluny 203645, and previously published individuals from Driffield Terrace9), different
ancient populations with connections to the Caucasus or the Pontic-Caspian region (Armenia_LBA, Armenia_Antiquity, Russia_
Sarmatian_Alan, and Russia_Sarmatian_PonticSteppe), and England_IA.

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Current Biology, Volume 34

Supplemental Information

An individual with Sarmatian-related

ancestry in Roman Britain
Marina Silva, Thomas Booth, Joanna Moore, Kyriaki Anastasiadou, Don
Walker, Alexandre Gilardet, Christopher Barrington, Monica Kelly, Mia
Williams, Michael Henderson, Alex Smith, David Bowsher, Janet
Montgomery, and Pontus Skoglund
Figure S1. Skeletal remains and aDNA authenticity, related to Figure 1.
A) Offord Cluny 203645/Burial 20.507 inhumation (Crick ancient genomics lab ID: C10271). B)
Frequency of misincorporations at the 5’- (left) and 3’- (right) ends of sequencing reads in the form of
C>T, denoting postmortem molecular damage (plot generated using DamageProfiler). Sequencing
metrics shown in Data S1.
Figure S2. Ancestry outlier Offord Cluny 203645, related to Figures 1 and 2.
A) Principal Component Analysis (PCA) shown in Figure 1C, with additional projected ancient
individuals (dated to between 2000-1475 BP) coloured according to geography. Offord Cluny 203645
is represented by a yellow square. Individuals included in the populations used as sources in the
qpWave/qpAdm models are grouped (as highlighted in Figure 1), with additional individuals from the
same regions coloured according to geographical location and data type (whole-genome shotgun
sequencing or ‘1240k’ SNP capture). Present day individuals are indicated by the first 3 letters of their
population label, as reported in Data S2E. B) Fixed qpAdm distal model using 3 sources: Western
European Hunter-gatherers (WHG), Balkan Neolithic (Balkan_N) and Steppe (Data S2F).
Transparency denotes the rejected model (p<0.05). C) 2-source qpAdm distal model with highest
p-value (p=0.551): Caucasus Hunter-gatherers (CHG) and Anatolia Neolithic (Anatolia_N) (Data
S2G). Individuals included in the models and population grouping listed in Data S2D.
Figure S3. f4-statistics, related to Figure 1.
Data points referring to Offord Cluny 203645 are shown in yellow, and data points for individuals from
Driffield Terrace site in York shown in grey. Error bars denote 1 standard error. See also Data S2A. A)
f4(South_Africa_400BP, England_IA; Roman individual, PopX). B) f4(South_Africa_400BP, Roman
individual; PopX, Russia_Sarmatian_PonticSteppe). C) f4(South_Africa_400BP, Roman individual;
Russia_Sarmatian_Alan, PopX).