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X/2/2019
INTERDISCIPLINARIA ARCHAEOLOGICA
NATURAL SCIENCES IN ARCHAEOLOGY
homepage: http://www.iansa.eu
Genetic Kinship and Sex Determination of Early Modern Period Human
Remains from a Defunct Graveyard in the Former Village of Obora
(Located on Šporkova Street in Prague’s Lesser Town District)
Jana Nováčková
a*
, Otakara Řebounová
b
,
Dana Kvítková
c
,
Martin Omelka
b
,
Vlastimil Stenzl
c
a
Institute of Archaeology CAS, Letenská 4, Prague, Czech Republic
b
Prague City Archives, Archivní 6, Prague, Czech Republic
c
Institute of Criminalistics, Bartolomějská 310/12, Prague, Czech Republic
1. Introduction
The implementation of genetic analyses into studies of
archaeological skeletal remains can provide information
about genetic kinship (Ciu
et al.
, 2015; Deguilloux
et al.
,
2014) and the genetic sex of children, when incomplete and
poorly-preserved skeletons (Álvarez-Sandova
et al.
, 2014;
Lassen
et al.
, 2000; Tierney, Bird, 2014) cannot be reliably
determined with diferent methods. Analyses of ancient
DNA (aDNA) have been previously used in demographic
studies of skeletal archaeological remains from several
archaeological sites in the Czech Republic, for example,
by Boberová
et al.
, 2012, Bravermanová
et al.
, 2018, or
Frolík
et al.
, 2017. The determination of genetic kinship
among the buried individuals would give an important
insight into understanding funerary practices, and the
social and demographic structures of historical cultures.
Additional useful information can also be obtained from
written historical sources, such as civil and parish registers,
testaments and chronicles.
The quality of genetic analyses of aDNA are negatively
infuenced by two major problems: its degradation into small
fragments; and the contamination of aDNA with modern
DNA. Firstly, over time, the DNA will become damaged
and broken into small fragments due to its inhospitable
environmental conditions (Hofreiter
et al.
, 2001,
pp. 353–354
;
Pääbo
et al.
, 2004, pp.654–660). Secondly, contaminant
DNA can come from individuals who were in contact
with the skeletal remains (archaeologists, anthropologists,
or geneticists in the laboratory), as well as from chemical
reagents, laboratory, or cross-sample contaminations. While
working with our samples for genetic analyses, we followed
the instructions published by Yang and Watt (2005).
Archaeogenetic research of genetic kinship is based
on analyses of uniparental markers (Y-chromosome
and mitochondrial DNA) and autosomal STR markers
Volume X ● Issue 2/2019 ● Pages 143–152
*Corresponding author. E-mail: novackova.janka@gmail.com
ARTICLE INFO
Article history:
Received: 1
st
May 2019
Accepted: 15
th
October 2019
DOI: http://dx.doi.org/ 10.24916/iansa.2019.2.4
Key words:
Early Modern Period
ancient DNA (aDNA)
genetic analyses
short tandem repeats
Y-chromosome
autosome
ABSTRACT
The main aim of this study was to determine genetic kinship and genetic sex of individuals buried
either in the same grave, multi-level grave, or neighbourhood grave. Success of genetic analyses is
based on the quantity and quality of extracted aDNA, which can be compromised by degradation
of DNA and possible contamination by modern DNA. We analysed archaeological skeletal remains
from an Early Modern period graveyard belonging to the Church of St. John the Baptist in the former
village of Obora, one of the most honourable Early Modern period archaeological sites in the Czech
Republic. Most of the 906 excavated anatomically-laid burials are dated to the years 1730s–1770s.
The results of 23 analysed individuals (divided into 4 groups) revealed that individuals are not blood
relatives. Studies of historical written sources provide information that the parish afliation at the time
of death had a crucial role in choosing the place for burial. Genetic analyses increased success rate of
sex determination to 91% compared to 61% determined by morphological methods. We were thus able
to determine the genetic sex of children, an evaluation that cannot be made by morphological methods.
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IANSA 2019 ● X/2 ● 143–152
Jana Nováčková, Otakara Řebounová, Dana Kvítková, Martin Omelka, Vlastimil Stenzl: Genetic Kinship and Sex Determination of Early Modern Period Human
Remains from a Defunct Graveyard in the Former Village of Obora (Located on Šporkova Street in Prague’s Lesser Town District)
144
(Deguilloux
et al.
, 2014; Juras
et al.
, 2017; Melchior
et al.
,
2010; Simón
et al.
, 2011). Each marker has its own unique
mechanism of heritability from parent to ofspring, and can
reveal or exclude genetic relationships at diferent levels. We
analysed the skeletal remains from a defunct graveyard of
the Church of St. John the Baptist in the former village of
Obora, situated at Šporkova Street no. 322/III in Prague, the
capital of the Czech Republic. Genetic kinship and genetic
sex was determined from the results of autosomal and
Y-chromosomal STR markers.
The site of Obora used to be a village located near Prague
castle in the quarter known as Prague’s Lesser Town. The frst
written record referencing Obora is dated to the years 1278–
1282, but previous excavation has uncovered fragments
dated to between the 9
th
–10
th
century (Dragoun, 1988a;
1988b; 1991). Obora was assigned to Prague in the 1650s,
and its Church of St. John the Baptist was incorporated into
the parish district of the Church of St. Wenceslas. The church
with its graveyard was closed in 1784, and rebuilt into a
residential building (Omelka, 2009). Skeletal remains of 906
anatomically-laid burials or parts thereof, that were dated to
the years 1730s–1770s according to their grave goods, were
excavated and documented during the archaeological rescue
excavation conducted by the Department of Archaeology of
the National Heritage Institute in Prague in the year 2002
(study no. 30/02) and 2004 (study no. 30/04) – Figure 1. The
archaeological location in Šporkova Street is one of the most
valuable Early Modern period archaeological sites in the
Czech Republic due to the assemblage collection of grave
goods and preserved written historical sources, providing
great possibilities to study: funerary customs among the
burgher citizens of the time (Omelka, Řebounová, 2017);
other manifestations of Baroque religiousness (Omelka,
Řebounová, 2011; 2014); as well as social and demographic
structures among this population (Omelka, Řebounová,
2012b). Several articles were published (mainly in Czech
peer-reviewed journals) regarding artefacts found in the
grave, including goods such as rings (Omelka, Šlancarová,
2007), beads (Omelka, Řebounová, 2008), crosses (Omelka
et al.,
2009; 2010), pins (Omelka
et al.,
2011), a medallion
(Omelka, 2006a; Omelka, Řebounová, 2012a; 2016) and
buttons (Omelka
et al.,
2018). Pilot results of genetic
analyses of 11 individuals were presented at the International
conference “Internationale Tagung der Österreichischen
Gesellschaft für Mittelalterarchäologie 2018” in Sankt
Pölten (Austria) (Nováčková
et al.,
in press). In the present
study, we increased the number of analysed individuals to
confrm or reject the hypothesis that the pattern of funerary
practices of Early Modern society, as suggested by the pilot
study, would hold up under further examination.
2. Material and methods
We analysed a total of 46 samples (bones and teeth) from
23 individuals (Table 1), of which 12 individuals (group 3 and
group 4) are newly published, and 11 individuals (group 1
and group 2) were previously published (Nováčková
et al.
,
in press). Individuals were divided into four groups (Table 1)
according to their stratigraphic relationships in the graveyard
(Figure 2). The groups contain the genetic material of men,
women and children, except for group 2, where two children
(newborn and 18 months old) were buried just above an
adult woman. Multi-level graves contained skeletal remains
of adult women, men and children and so there is a high
probability that they are members of one family (for example,
Figure 1.
The plan of three phases of
excavations at the defunct graveyard of the
Church of St. John the Baptist and ground
plan of the church. The archaeological
rescue excavation was carried out only on
the part of graveyard in which construction
work took place on (Omelka, 2006b,
unpublished). Drawn by Martin Omelka.
0 20 m
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IANSA 2019 ● X/2 ● 143–152
Jana Nováčková, Otakara Řebounová, Dana Kvítková, Martin Omelka, Vlastimil Stenzl: Genetic Kinship and Sex Determination of Early Modern Period Human
Remains from a Defunct Graveyard in the Former Village of Obora (Located on Šporkova Street in Prague’s Lesser Town District)
145
Figure 2.
Flowcharts by Jiří Vachuda.
The fowcharts are parts of unpublished
documentation of research in Šporkova
Street, schematically representing the
position of graves on the burial site in the
geometrically defned sectors. Each sector
is designated by a diferent colour. Some
skeletons intersected more than one sector,
and so are labelled using more than one
colour. The results of the computer analysis
of genetic kinship are the nearby fowcharts.
Figure 3.
Skeletal remains of individuals
H66 and H71 (in blue). Grave H66 contained
skeletal remains of an adult female with
additional bones (in green) of a child and
adult male. Photographed by Jiří Vachuda.
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IANSA 2019 ● X/2 ● 143–152
Jana Nováčková, Otakara Řebounová, Dana Kvítková, Martin Omelka, Vlastimil Stenzl: Genetic Kinship and Sex Determination of Early Modern Period Human
Remains from a Defunct Graveyard in the Former Village of Obora (Located on Šporkova Street in Prague’s Lesser Town District)
146
Table 1.
The list of individuals analysed.
No.IDSex determined
anthropologically
Age determined
anthropologically
Analysed part of
skeleton
Concentration
of aDNA (pg/ul)
Inventory
number
Group
1.
H72/I
female
30–40
tooth
33.4
P7A 19 001
1
2.
H72/II
bone
4.64
3.
H78/I
undetermined
2–3
tooth
1.55
P7A 19 007
1
4.
H78/II
bone
36
5.
H94/I
undetermined
juvenis
tooth
4.22
P7A 19 021
1
6.
H94/II
bone
5.25
7.
H96/I
female
50–60
metatarsal
0.24
P7A 19 023
1
8.
H96/II
rib
8.03
9.
H98/I
female
30–40
tooth
2.46P7A 19 025
1
10.
H196/I
malematurus
tooth
15.3
P7A 19 122
1
11.
H196/II
rib
11.6
12.
H222/I
male
40–50
ulna
394
P7A 19 162
1
13.
H222/II
tooth
366
14.
H251/I
female
30–40
tooth
2.11
P7A 19 173
1
15.
H251/II
tooth
0.9
16.
H120/I
undeterminednewborn
ulna
9.06
P7A 19 047
1
17.
H120/II
rib
66
18.
H242/I
undetermined
juvenis
bone
4.32
P7A 19 162
1
19.
H242/II
rib
2.12
20.
H64/I
undetermined18 months
tooth
1.26
P7A 18 993
2
21.
H64/II
bone
2.54
22.
H65/I
undeterminednewborn
humerus
35.7
P7A 18 994
2
23.
H65/II
rib
68.1
24.
H101/I
femalematurus
tooth
47.4
P7A 19 028
2
25.
H101/II
rib
1.25
26.
H66a/I
femalematurus
tooth
6
P7A 18 995
3
27.
H66a/II
tooth
17.8
28.
H66b/I
undetermined
juvenis
rib
11.4
29.
H66b/II
bone
2.04
30.
H66c/I
malematurus
tooth
2.79
31.
H66c/II
tooth
31.3
32.
H71/I
undeterminednewborn
femur
2.62
P7A 19 000
3
33.
H71/II
os petrosum
69.2
34.
H85/I
female
20–30
tooth
0.5
P7A 19 014
3
35.
H85/II
tooth
5.95
36.
H85/III
tooth
2.16
37.
H82/I
undeterminednewborn
radium
3.8
P7A 19 011
4
38.
H82/II
phalang
2.93
39.
H97/I
female
50–60
calf bone
2.49
P7A 19 024
4
40.
H97/II
phalang
1.29
41.
H165/I
female
40–50
tooth
35.2
P7A 19 092
4
42.
H165/II
os petrosum
396
43.
H145/I
maleadultus
os petrosum317
P7A 17 072
4
44.
H145/II
tooth
17.5
45.
H192/I
maleadultus
tooth
24.9
P7A 19 118
4
46.
H192/II
metacarpus
1.97
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Jana Nováčková, Otakara Řebounová, Dana Kvítková, Martin Omelka, Vlastimil Stenzl: Genetic Kinship and Sex Determination of Early Modern Period Human
Remains from a Defunct Graveyard in the Former Village of Obora (Located on Šporkova Street in Prague’s Lesser Town District)
147
see Figure 3). Samples of teeth and bones were taken from
diferent parts of the skeletons, depending on their state of
skeletal preservation. For detailed information about the
samplings, see Table 1. Sampling took place in the National
Museum in Prague, where the remains are deposited.
Samples were analysed in several independent steps in
four separate rooms (mechanical cleaning; extraction of
aDNA; quantifcation and PCR amplifcation; post-PCR
sequencing). Blank controls were added to each step/reaction
to monitor for possible contamination resulting from the lab
procedures, but revealed no evidence thereof.
2.1 Mechanical cleaning and extraction of aDNA
Samples were rinsed using 96% ethanol and ultra clean
water. Bone and teeth surfaces were sanded using either a
Dremel Multi (Dremel) electric mini sander, or manually
using sandpaper due to a sample’s preservation. Bones
were cut into small pieces and ground into powder using a
6870 Freezer Mill (Spex Sample Prep). Subsequently, 70mg
of the bone powder was incubated in a lysis bufer (0.5M
EDTA [pH 8.0], Proteinase K and 0.5% SDS) at 56°C in a
UVP HB –1000 Hybridizer Hybridization Oven (Analytik
Jena US LLC) for 24 hours. Finally, aDNA was extracted
using a MinElute PCR Purifcation Kit (Qiagen) according
to a modifed protocol published by Yang
et al.
(1998)
and Anderung
et al.
(2008).
2.2 Quantifcation of aDNA
The success of the extraction of preserved aDNA and the
amount of extracted aDNA was determined using a real-time
PCR quantifcation Plexor HY System (Promega) kit on a
LightCycler 480 RealTime PCR Instrument (Roche). Samples
were prepared in duplex reactions. The Plexor HY kit contains
primer for a target of a 133bp sequence from a testis-specifc
protein, Y-encoded (TSPY) locus on chromosome Y, providing
means of determining genetic sex.
2.3 Amplifcation and sequencing of aDNA
Samples were analysed for 23 autosomal STR markers,
amelogenin X and Y, and 23 Y-chromosomal STR markers
using four commercially-available kits: the PowerPlex ESX
17 System, the PowerPlex ESI 17 Pro System, the PowerPlex
16 System, and the PowerPlex Y23 System (all from Promega
Corporation). Amelogenin X and Y loci were used to
determine genetic sex. Each sample was analysed in several
independent amplifcations and sequencing reactions using
peqSTAR 96X Universal Gradient cycler (VWR Peqlab)
and the Applied Biosystems 3130xl Genetic Analyzers
instrument (15kV injection for 15s at POP4 polymer)
(Applied Biosystems). Samples were prepared according to
the manufacturer’s recommendation with 32 amplifcation
cycles instead of the 30 that were recommended by the
manufacture protocol.
2.4 Data analyses
Raw data from capillary electrophoresis were analysed with
GeneMapper IDX software (Applied Biosystems). Results
of autosomal and Y-chromosomal STR markers were used
for genetic kinship and genetic sex determination among
buried individuals. Results of STR markers were evaluated
and computed by several software programs: Mlrelate
(Kalinowski
et al.
, 2006), Familias 3 (Kling
et al.
, 2014),
Network 5 (Bandelt
et al.
, 1999), and Network Publisher
2.1.2.5 (Fluxus Technology LtD.) A phylogenetic network
of Y-chromosomal STR markers was constructed only
for markers that were successfully genotyped in all male
samples. The network was constructed using Median joining
(Bandelt
et al.
, 1999), and the fnal tree was redrawn by
Network Publisher 2.1.2.5 (Fluxus Technology LtD). Genetic
kinship between samples in all groups was computed by
one to one for all samples using software Familias 3, that
diferentiated between fve categories of genetic relationship
(parent-ofspring, full siblings, half siblings, cousins and
second cousins) as well as unrelated individuals using Blind
search by calculating a likelihood ratio (Kling
et al.
, 2014)
and ML-Relate (Kalinowski
et al.
, 2006) that determined
three close relationships (parent-ofspring, full siblings and
half siblings) and unrelated individuals (Kalinowski
et al.
,
2006).
3. Results
The success of the genetic analyses depends on the quantity
and quality of the extracted aDNA (Table 1). Genetic sex
and genetic kinship was evaluated for individuals that were
successfully genotyped in several independent reactions of
two or three diferent samples from one individual. Samples
H64 and H94 were excluded from the statistical analyses,
since sample H64 failed to be successfully genotyped, and
sample H94 did not provide reliable results (results from two
diferent parts of the skeleton gave diferent results). The
skeletal remains of H94 were excavated during two diferent
archaeological excavations and it is possible that the two
parts of the skeleton were completed incorrectly.
3.1 Genetic kinship
Genetic kinship was determined from the result of autosomal
(Table 2) and Y chromosomal STR markers (Table 3). STR
profles listed in Table 2 and Table 3 are summaries of all
performed analyses from all kits, as well as all samples
analysed for the same individual. The success rate of
STR marker detection was increased using three diferent
available autosomal kits. Ancient DNA is degraded into
small fragments over time, and it is necessary to analyse
small fragments (Allentoft
et al.
, 2012; Pääbo, 1989).
The advantage of using the PowerPlex ESX 17 System
and PowerPlex ESI 17 Pro System kits as complements
is that while they contain the same markers, the primers
are designed to complement one another, with a diferent
fnal marker length. Markers that are long in the frst kit
are short in the second. The stratigraphic relation between
individuals within the burial grounds is shown on fowcharts
below (Figure 2). Genetic analyses revealed only unrelated
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Jana Nováčková, Otakara Řebounová, Dana Kvítková, Martin Omelka, Vlastimil Stenzl: Genetic Kinship and Sex Determination of Early Modern Period Human
Remains from a Defunct Graveyard in the Former Village of Obora (Located on Šporkova Street in Prague’s Lesser Town District)
148
Table 2.
Results of autosomal STR markers (NA – results not available; XX – female; XY – male).
Amel
D3S1358
D19S433
D2S1338
D22S1045
D16S539
D18S51
D1S1656
D10S1248
D2S441
TH01
vWa
D21S11
D12S391
D8S1179
FGA
SE33
Penta E
D5S818
D13S317
D7S820
CSF1PO
Penta D
TPOX
H72
XX
16; 17
15; 13
16; 17
1512; 1315; 17
12; 17.3
1414
8; 916; 19
30
17; 19
12; 1423; 2514; 2112
9; 11
8; 121111
9
11
H78
XX
14; 16
13; 15
NA
151113; 14
12; 15.313; 16
10
6; 8
17; 18281514; 1524
NANA
11; 1210; 128; 11
NANA9; 11
H94
NA
H96
XX15; 1813
NA11; 16NANA11; 16
14107; 8
16; 17
28
19; 24
11
NANANA
1311
NANANA
8
H98
XX14; 171320; 25
15; 169; 11
13; 17
12; 16
1510; 11
7; 915; 19NA15; 19
12; 1320; 2417131114
9NANA
8
H196
XY15; 17 1524
16
12; 1212; 14
13; 17.3
1411; 117; 8
16; 1829; 30
22; 24
13; 1619; 2620; 27.2NA
11; 1211; 1211; 1212
NA8; 9
H222
XY
14; 16
13; 14
17; 19
15; 181112; 14
16
13; 1510; 127; 717; 18
28; 29
18; 2412; 1321; 22
19.2; 21.2NA
11810; 1212
NA9
H251
XX
15; 16
12
16
15121812; 1414
10; 11.38; 9.3
17; 1828; 30
NA
13; 1422; 25
25.2; 17NA
118; 11
NANANANA
H242
XX14; 171217; 18
14; 16
11; 1212; 1411; 151410; 11
9; 9.3
20
30; 30.2
20; 22
NANA
18
NANANA
1011
NANA
H64
NA
H65
XX
16; 17
1423
169; 13
1212; 151311
6; 7
18
29; 31
18; 2011; 1223; 252071111; 121010
NA
10; 11
H120
XX15; 1814; 152014; 1511; 1211; 121115; 1711; 14
8; 9.3
15; 18
31.218; 19
12; 1320
23.2; 25.2NA
10; 131211
NANA
8; 12
H101
XX18
14; 15.2
17
12; 16
1115; 18
15.3; 18.3
1314
6; 9.31629; 32.2
2214
19; 2024.2; 30.2
10; 1811; 121110138; 1211
H66a
XX15; 1711; 1323; 24
169; 11
12; 1411; 12
13; 1611; 11.36; 9
14; 17
28; 2917; 19
1022; 23
18; 27.211; 16NA
11; 121110108; 11
H66b
XX15; 1813
NA
11; 17
9; 12
15
13; 17.316
10
6; 9.314; 19
28; 311811
NANA
13111211
NANA
8; 14
H66c
XY
13; 16
15; 171712; 1711; 1215; 17
18.3
13; 1410; 11
6; 816; 1827; 32.2
1813; 15
20.2; 2321.2; 30.2NANANANANANANA
H71
XX
16; 17
1517; 2011; 1511; 1312; 14
12; 16
13; 1411
6; 9.316; 1828; 30.2
15; 1814; 1525
NA
5; 15
11.12
8
8; 1012
9; 12
8; 12
H85
XX14; 17
13; 14.2
23; 2411; 178; 1312; 1412; 1712; 1311
7; 9.3
14; 17
29; 30
20; 2110; 14
19; 2515; 19
5
9; 13
8; 138; 1213
9; 13
8
H82
XY
16; 1814; 16.2
17; 23
15; 16
11; 1313; 1411; 151414
NA
17; 1828; 301510; 1422; 23
16; 25.2NA
10; 128; 11
8; 9
10
NA
8
H97
XX1512; 1317
16; 17
12
NANA16
147
16NANA
15; 1723; 24
NANA
1111; 1410; 1212
NANA
H165
XY14; 15
14; 15.2
23; 251111; 1415; 17
16
13; 1711; 14
9; 9.315; 1629; 30
17; 201321; 23
25.2; 28.2
5; 121210; 121010118
H145
XX14; 1511; 14231510; 1211; 13
15; 16
12; 1310; 14
6; 9.316; 1730; 32.2
1812; 15
23; 23.226.2; 27.2
7
9; 12
1110
NANA
11
H192
XY
16; 2014; 15.218; 1916; 17
1213; 15
15; 15.3
13; 1510; 11
9; 9.3
15; 18
29; 3016; 17
13; 1521; 23
16; 20NA
10; 1311; 1211; 1211
9; 138; 9
Table 3.
Results of Y-chromosomal STR markers (NA – results not available).
DYS576
DYS389I
DYS448
DYS389II
DYS19
DYS391
DYS481
DYS549
DYS533
DYS438
DYS437
DYS570
DYS635
DYS390
DYS439
DYS392
DYS643
DYS393
DYS458
DYS385a
DYS385b
DYS456
YGATA–H4
H196
18132032
NA
1130111210
NA
18242412111013
16
14
16
15
NA
H222
17131828141222121212151823221113
NA
13181114
16
12
H66c
1713
19
30
NA
10221212
NANA
1724
NANANANA
13
16
1114
NANA
H82
1813
1929
14
9
2312
NANANA
1720
NA
1211
9
12
19
1315
NANA
H165
16
1320301410
26
121010
16
212123111112131714151411
H192
1813203014102212121014
19
21241311
NA
13
1616
181711
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Jana Nováčková, Otakara Řebounová, Dana Kvítková, Martin Omelka, Vlastimil Stenzl: Genetic Kinship and Sex Determination of Early Modern Period Human
Remains from a Defunct Graveyard in the Former Village of Obora (Located on Šporkova Street in Prague’s Lesser Town District)
149
relationships between analysed samples within all groups.
The matrix generated by Ml-Relate software (Figure 2),
provides information that individuals in all groups are not
blood-related.
Six individuals were determined by a signal for amelogenin
Y as a male and were analysed for Y-chromosomal STR
markers (Table 3). All identifed male individuals difered
considerably from each other in terms of observed alleles
(Figure 4), providing no evidence of any father-son
relationships, nor of a common close male ancestor.
Figure 4.
The phylogenetic network constructed for Y-chromosomal STR
markers (yellow rings-individuals; red numbers-number of mutations).
Table 4.
Results of morphological and genetic sex determination; discordances between morphological and genetic fndings are labelled in red.
No.IDSex determined
antropologically
Sex determined
genetically
No.IDSex determined
antropologically
Sex determined
genetically
1.
H72
femalefemale
13.
H101
femalefemale
2.
H78
undeterminedfemale
14.
H66a
femalefemale
3.
H94
undeterminedundetermined
15.
H66b
undeterminedfemale
4.
H96
femalefemale
16.
H66c
malemale
5.
H98
femalefemale
17.
H71
undeterminedfemale
6.
H196
malemale
18.
H85
femalefemale
7.
H222
malemale
19.
H82
undeterminedmale
8.
H251
femalefemale
20.
H97
femalefemale
9.
H120
undeterminedfemale
21.
H165
femalemale
10.
H242
undeterminedfemale
22.
H145
malefemale
11.
H64
undeterminedundetermined
23.
H192
malemale
12.
H65
undeterminedfemale
3.2 Sex determination
The presence/absence of a signal for amelogenin Y locus
was used to determine the genetic sex of skeletal remains
(XX – female and XY – male). Six individuals determined
as a male by the presence of a signal for amelogenin Y were
successfully genotyped for Y-chromosomal STR markers.
Due to the fact that the amelogenin Y locus can be afected by
allelic drop-out, we also took the result of the amplifcation
TSPY gene in the Plexor HY kit into consideration. All
samples that did not have a signal for amelogenin Y were
also not amplifed for the Y-chromosomal TSPY gene,
and genetic sex was classifed as a female (Table 4). Our
results were compared with the morphological fndings of
studies performed by Milan Stloukal from the Department
of Archaeology of the National Museum in Prague. His
unpublished morphological examinations of skeletal remains
are archived in the Department of Archaeology in the
National Heritage Institute in Prague. The anthropological
sex and age of the skeletal remains were determined using
methods in accordance to the protocol by Ferembach
et al.
(1979). We observed two cases of discordance (for
individuals H145 and H165) between morphological and
genetic fndings (see Table 4), and thus were able to increase
the rate of sex determination from 61% for morphological
fndings (14 individuals) to 91% (21 individuals) for genetic
fndings. We were also able to determine the sex of children
that could not be evaluated by morphological methods. Both
individuals H145 and H165 were poorly preserved, having
seriously damaged skeletons and fragmented skulls.
4. Discussion
The results of the genetic analyses confrmed the hypothesis
about the funerary practices of Early Modern period
burghers, which was based on the study of historical
written sources such as death registers, parish registers and
testaments. Historical written sources did not provide any
clear information about the existence of family graves on the
bourgeois graveyard of St. John the Baptist church in the
in Early Modern period village of Obora. Genetic analyses
revealed that the individuals, who were buried in the same
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Jana Nováčková, Otakara Řebounová, Dana Kvítková, Martin Omelka, Vlastimil Stenzl: Genetic Kinship and Sex Determination of Early Modern Period Human
Remains from a Defunct Graveyard in the Former Village of Obora (Located on Šporkova Street in Prague’s Lesser Town District)
150
multi-level graves or in neighbouring graves, are not blood-
related members of a single family.
The evidence from parish and civil registers suggests that
bourgeois (middle class) members of society were buried in
accordance to the parish afliation of the given house which
was the site of their death. Division of a family at the time
of death was not unusual in that period. As a case in point,
we can mention the burials of the young boy Johann Roßlaw
and his mother. Although the boy died on 3
rd
May 1764 and
his mother on 22
nd
May 1764, they were not buried in the
same grave, nor even at the same graveyard as a result of the
diferent parish afliation of the houses in which their death
took place. The boy died at the “House of Three Swallows”
and was buried at the graveyard of the Church of St. John
the Baptist, but his mother died at “Schumann House”, and
was buried at the graveyard of the Church of St. Lawrence.
Her daughter Rosina died in October 1764 at the house “U
Jedličků” and was buried at the graveyard of the Church of the
St. Lawrence (Prague City Archives, Collection of Matrices,
sign. MIK Z4, fold 19–20 and 22; Omelka, Řebounová,
2012a, p.239). A very similar example was in the case of
the Roßenfeld family. Three of the fve children were buried
at the graveyard of the Church of St. Lawrence and two at
the graveyard of the Church of St. John the Baptist. It was
also probably due to their diferent parish afliations (Prague
City Archives, Collection of Matrices, sign MIK Z3, fold
310 and 313; sign MIK Z4, fold 20 and 31).
Other useful sources of funerary practices can be gleaned
from testaments. Testaments of aristocratic and the richest,
social-bourgeoisie class individuals, who were usually buried
in church interiors, were written very precisely, containing
detailed information about where exactly they want to be
buried and even with whom they wish to be buried after
death (Král, 2005; Nováčková
et al.,
2019, in press). The
graveyard of the Church of St. John the Baptist was mainly
used to bury citizens belonging to the middle or lower-middle
social classes, and individual testaments (if written) usually
only specifed the name of the church. We can mention,
for example, the testament of František Dispach. He died
in 1766 in his house in Lesser Town, which belongs to the
parish district of the Church of St. John the Baptist. His
testament was written only several months before his death,
and he wanted to be buried at the graveyard of the Church
of St. John the Baptist or the graveyard of the Church of St.
Wenceslas. Finally, he was buried at the graveyard of the
Church of St. Wenceslas (Manuscript Collection, sign 4764,
fol. A21–A22) and not at the graveyard of the Church of St.
John the Baptist, where his fve children had been buried
before him (Anna died in 1731, Vaclav in 1737, Theresie
in 1743, Ludmila in 1743, and Antonie in 1750). All his
children died in their father’s house (Prague City Archives,
Collection of Registries, sign MIK 3, fol. 185, 211, 253,
256 and 281). There is also mention of Anna Dispachova,
who died in 1742 in the house “At the White Angel”, and
was buried at the graveyard of the Church of St John the
Baptist, but the relationship with František Dispacha is
not clear from the register (Archive of the City of Prague,
Collection of Registries, sign MIK 3, fol. 236). It is evident
that family relationships were not taken into consideration
when members of one family were buried.
According to the data from historical written sources and
from the results of genetic analyses, there is no indication
that people from the Early Modern period’s lower and middle
social classes (
i.e.
most of the people buried at graveyard
of the Church of the St. John the Baptist) of purposefully-
buried members of one family were buried in one multi-
level grave, or in neighbourhood grave sites, or even in
the same graveyard, as was the very common practice of
the aristocracy and the richest among the population (Král,
2005).
5. Conclusion
In this study we have applied an interdisciplinary approach
to investigate genetic kinship, genetic sex and the funerary
practices of an Early Modern period bourgeois society.
Genetic analyses are a powerful method for sex determination
in skeletons of children, as well as in badly-preserved and
incomplete skeletons of adults (Álvarez-Sandova
et al.
, 2014;
Lassen
et al.
, 2000; Tierney, Bird, 2014), where morphological
methods provide unreliable or no results (Álvarez-Sandova
et al.
, 2014; Lassen
et al.
, 2000; Tierney, Bird, 2014). We
observed a contradiction between morphological and genetic
methods in the sex determination of two separate skeletons
of buried individuals: individuals H145 and H165 were
poorly preserved, resulting in an unreliable morphological
determination of sex. In such cases, genetic analyses are a
more exact method to determine the sex of skeletal remains
than are morphological treatments. By implementing genetic
analyses, the number of successfully-determined individuals
increased from 14 (61%) to 21 (91%); in addition, we were
able to determine the genetic sex of children that could not
be determined through morphological methods.
Genetic analyses are a crucial tool in determining the
genetic kinship of archaeological skeletal remains. The
skeletal samples used for genetic analyses were chosen
according to their relative stratigraphic positions within the
burial grounds, and divided into four groups. Skeletons of
adults and children buried in the same multi-level grave,
or in very close proximity, have a high probability of being
members of the same family. Genetic analyses of autosomal
and Y-chromosomal STR markers revealed, however, that the
individuals analysed were not blood-relatives. These results
of genetic analyses are in accordance with and confrm
the hypothesis based on the evidence provided by written
historical sources (civil and parish registers and testaments).
There is evidence that some members of families of middle
and lower social classes were buried in diferent graveyards:
because they had died in diferent houses belonging to a
diferent parish afliation. The tradition of founding family
graves at that time is well documented among aristocratic
families and the more wealthy inhabitants, who were usually
buried together; however, this would be in the interior of the
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IANSA 2019 ● X/2 ● 143–152
Jana Nováčková, Otakara Řebounová, Dana Kvítková, Martin Omelka, Vlastimil Stenzl: Genetic Kinship and Sex Determination of Early Modern Period Human
Remains from a Defunct Graveyard in the Former Village of Obora (Located on Šporkova Street in Prague’s Lesser Town District)
151
church. On the other hand, the majority of baroque inhabitants
of the past village of Obora buried at the graveyard of the
Church of St. John the Baptist were probably buried there
wherever a free place was available and according to their
parish afliation, without taking blood relationships into
consideration.
Acknowledgements
The research of family relationships of skeletal remains was
supported by a grant from the Ministry of the Interior of the
Czech Republic (VI20162020015). We are grateful to the
National Museum in Prague for allowing us to sample the
human remains in their possession.
References
ALLENTOFT, M.E., COLLINS, M., HARKER, D., HAILE, J., OSKAM,
C.L., HALE, M.L., CAMPOS, P F., SAMANIEGO, J.A., GILBERT,
M.T., WILLERSLEV, E, ZHANG, G., SCOFIELD, R P., HOLDAWAY,
R.N., BUNCE, M., 2012. The half-life of DNA in bone: measuring
decay kinetics in 158 dated fossils.
Proceedings Biological Sciences
,
279(1748), 4724–4733.
ÁLVAREZ-SANDOVAL, B.A., MANZANILLA, L.R., MONTIEL, R.,
2014. Sex Determination in Highly Fragmented Human DNA by High-
Resolution Melting (HRM) Analysis.
PLoS ONE
, 9(8), e104629.
ANDERUNG, C., PERSSON, P., BOUWMAN, A., ELBURG, R.,
GÖTHERSTRÖM, A., 2008. Fishing for ancient DNA.
Forensic Science
International: Genetics
, 2(2), 104–107.
BANDELT, H.J., FORSTER, P., RÖHL, A., 1999. Median-joining networks
for inferring intraspecifc phylogenies.
Molecular Biology and Evolution
,
16(1), 37–48.
BOBEROVÁ, K., DROZDOVÁ, E., PÍŽOVÁ, K., 2012. Application of
Molecular Genetic Methods in Anthropological and Paleodemographic
Studies of Fragmentary and Damaged Skeletal Material from Rescue
Excavations.
Journal of Life Sciences
, 6(9), 961–969.
BRAVERMANOVÁ, M., DOBISÍKOVÁ, M., FROLÍK, J., KAUPOVÁ,
S., STRÁNSKÁ, P., SVĚTLÍK, I., VANĚK, D., VELEMÍNSKÝ, P.,
VOTRUBOVÁ, J. 2018. Nové poznatky o ostatcích z hrobů K1 a K2
rotundy sv. Víta na Pražském hradě – New fndings on the remains
from graves K1 and K2 from the St. Vitus Rotunda at Prague Castle.
Archeologické rozhledy
, 70, 260–293.
CUI, Y., SONG, L., WEI, D., PANG, Y., WANG, N., LI, C., FENG, B.,
TANG, W., LI, H., REN, Y., ZHANG, C., HUANG, Y., HU, Y., ZHOU,
H., 2015. Identifcation of kinship and occupant status in Mongolian
noble burials of the Yuan Dynasty through a multidisciplinary approach.
Philosophical Transactions of the Royal Society B: Biological Sciences
,
370(1660), 20130378.
DAMGAARD, P.B., MARGARYAN, A., SCHROEDER, H., ORLANDO,
L., WILLERSLEV, E., ALLENTOFT, M.E., 2015. Improving access to
endogenous DNA in ancient bones and teeth.
Scientifc Reports
, 5, 11184.
DEGUILLOUX, M., PEMONGE, M., MENDISCO, F., THIBON, D.,
CARTRON, I., CASTEX, D., 2014. Ancient DNA and kinship analysis
of human remains deposited in Merovingian necropolis sarcophagi (Jau
Dignac et Loirac, France, 7
th
–8
th
century AD).
Journal of Archaeological
Science
, 41, 399–405.
DRAGOUN, Z., 1988a. Archeologický výzkum rotundy sv. Jana Křtitele
pod Pražským hradem v r. 1986 a 1987 – Archaeological Excavation of
the Rotunda of St. John the Baptist below Prague Castle in 1986–1987 (in
Czech).
Archaeologia historica
, 13, 403–416.
DRAGOUN, Z., 1988b. Praha 1 – Malá Strana, Jánský vršek, Šporkova
ulice – Prague 1 – Lesser Town, Jánský vršek, Šporkova street (in Czech).
Pražský sborník historický
, 21, 184–185.
DRAGOUN, Z. 1991. Praha 1 – Malá Strana, Jánský vršek – Prague 1 –
Lesser Town, Jánský vršek, Šporkova street (in Czech).
Pražský sborník
historický
, 24, 195–196.
FEREMBACH, D., SCHWIDETZKY, I., STLOUKAL, M., 1979.
Empfehlungen fur die Alters- und Geschlechtsdiagnose am Skelett.
Homo
, 30, 1–32.
FROLÍK, J., STRÁNSKÁ, P., VOTRUBOVÁ, J., EMMEROVÁ, B.,
VANĚK, D., 2017. People “on the Margin”: A Medieval Cemetery
in Český Brod – Malechov (Central Bohemia).
Interdisciplinaria
Archaelogica: Natural Sciences in Archaeology
, 8(1), 59–75.
GAMBA, C., JONES, E.P., TEASDALE, M.D., McLAUGHLIN, R.L.,
GONZALES-FORTES, G., MATTIANGELI, V., DOMBRÓCZKI, L.,
KŐVÁRI, I., PAP, I., ANDERS, A., WHITTLE, A., DANI, J., RACZKY,
P., HIGHAM, T.F.G., HOFREITER, M., BRADLEY, D.G., PINHASI, R.,
2014. Genome fux and stasis in a fve millennium transect of European
prehistory.
Nature Communications
, 5(5257), 5257.
HANSEN, H.B., DAMGAARD, P.B., MARGARYAN, A., STENDERUP,
J., LYNNERUP, N., WILLERSLEV, E., ALLENTOFT, M.E., 2017.
Comparing Ancient DNA Preservation in Petrous Bone and Tooth
Cementum.
PLoS ONE
. 12(1), e0170940.
HOFREITER, M., SERRE, D., POINAR, H.N., KUCH, M., PÄÄBO, S.,
2001. Ancient DNA.
Nature Reviews Genetics
, 2(5), 353–359.
JURAS, A., CHYLENSKI, M., KRENZ-NIEDBALA, M., MALMSTROM,
H., EHLER, E., POSPIESZNY, L., LUKASIK, S., BEDNARCZYK, J.,
PIONTEK, J., JAKOBSSON, M., DABERT, M., 2017. Investigating
kinship on Neolithic post-LBK human remains from Krusza Zamkova,
Poland using ancient DNA.
Forensic Science International: Genetics
,
26, 30–39.
KALINOWSKI, S.T., WAGNER, A.P., TAPER, M.L., 2006. ML-Relate:
a computer program for maximum likelihood estimation of relatedness
and relationship. Molecular Ecology Notes, 6(2), 576–579.
KEYSER-TRACQUI, C., LUDES, B., 2005. Methods for the study of
ancient DNA.
Methods in molecular biology
, 297, 253–264.
KLING, D., TILLMAR, A.O., EGELAND T., 2014. Familias 3-Extensions
and new functionality.
Forensic Science International: Genetics
, 13,
121–127.
KRÁL, P. 2005: Tod, Begräbnisse und Gräber. Funeral ritus des böhmischen
Adels als Mittel der Representation und des Andenkens. In: M. Hengere,
ed.
Macht und Memoria. Begräbniskultur europäischer Oberschichten in
der Frühen Neuzeit.
Köln, Weimar, Wien: Böhlau, pp. 421–448.
LASSEN, C., HUMMEL, S., HERRMANN, B., 2000. Molecular sex
identifcation of stillborn and neonate individuals (“Traufkinder”) from
the burial site Aegerten.
Anthropologischer Anzeiger
, 58(1), 1–8.
MELCHIOR, L., LYNNERUP, N., SIEGISMUND, H.R., KIVISILD, T.,
DISSING, J., 2010. Genetic Diversity among Ancient Nordic Populations.
PLoS One
, 5(7), e11898.
NOVÁČKOVÁ, J., OMELKA, M., ŘEBOUNOVÁ, O., STENZL, V.,
in press. Begräbnispraxis des Pragerbarocken Bürgertums im Licht
der DNA-Analyse. In:
Internationale Tagung der Österreichischen
Gesellschaft für Mittelalterarchäologie 2018
. Sankt Pölten. Wien:
Beiträge zur Mittelalterarchäologie in Österreich.
OMELKA, M., 2006a. Nález neobvykle členěného medailonu s vyobrazením
kříže svatého Benedikta a Zachariášova požehnaní ze Šporkovy ulice čp.
332/III v Praze – The Finds of Unusually Divided Medallion with a Motif
of St. Benedict’s cross and Zachary’s Blessing from Šporkova Street, House
No. 322/III in Prague (in Czech).
Archeologica Pragensia
, 18, 144–152.
OMELKA, M., 2006b.
Investorská zpráva o archeologickém výzkumu
Praha 1 – Malá Strana, Šporkova ulice čp. 332/III – Investor’s report
about archaeological research in Prague 1–The Lesser Town, Šporkova
Street no. 322/III.
Unpublished manuscript deposited in the Archive of the
National Heritage Institute, ú. o. p. in Prague, Archive of the Department
of Archaeology.
OMELKA, M., 2009. Hřbitov u kostela sv. Jana v Oboře ve Šporkově ulici
čp. 332/III na Malé Straně v Praze – The Cemetery of the Church of St.
John in Šporkova Street (no.322/III), the Lesser Town, Prague (in Czech).
Staletá Praha
, 25, 93–101.
OMELKA, M., PETŘÍK, J., PROKEŠ, L., ŘEBOUNOVÁ, O.,
ŠLANCAROVÁ, V., 2018. Soubor knofíků ze zaniklého hřbitova při
kostelu sv. Jana v Oboře (Praha – Malá Strana) – An Assemblage of
Buttons from the Defunct Cemetery at the church of Saint John the Baptist
in Obora (Prague Lesser Town) (in Czech).
Archeologie ve středních
Čechách
, 22, 709–744.
image/svg+xml
IANSA 2019 ● X/2 ● 143–152
Jana Nováčková, Otakara Řebounová, Dana Kvítková, Martin Omelka, Vlastimil Stenzl: Genetic Kinship and Sex Determination of Early Modern Period Human
Remains from a Defunct Graveyard in the Former Village of Obora (Located on Šporkova Street in Prague’s Lesser Town District)
152
OMELKA, M., ŠLANCAROVÁ, V., 2007. Soubor prstenů ze zaniklého
hřbitova při kostelu sv. Jana v Oboře (Praha – Malá Strana) – A Collection
of Rings from the Defunct Cemetery at the church of Saint John the
Baptist in Obora, Prague – the Lesser Town (in Czech).
Archeologie
ve středních Čechách
, 11, 671–709.
OMELKA, M., ŘEBOUNOVÁ, O., 2008. Soubor korálků ze zaniklého
hřbitova při kostelu sv. Jana v Oboře (Praha1 – Malá Strana) – A
Collection of Beads from the Defunct Cemetery at the Church of St. John
the Baptist in Obora, Prague–the Lesser Town (in Czech).
Archeologie
ve středních Čechách
, 12, 606–679.
OMELKA, M., ŘEBOUNOVÁ, O., 2011. Poznámky ke zbožnosti
a pohřebnímu ritu malostranského barokního měšťanstva ve světle
archeologických nálezů – Remarks on the Religiousness and Burial Rite
of the Lesser Town Baroque Burghers in the Light of Archaeological
Findings (in Czech).
Pražský sborník historický
, 39, 268–298.
OMELKA, M., ŘEBOUNOVÁ, O., 2012a. Soubor medailonů a feniků
se symbolikou sv. Benedikta ze zaniklého hřbitova při kostelu sv. Jana
v Oboře (Praha1 – Malá Strana) – A Collection of Medallions and
Pfennigs with the Symbols of St. Benedict from the Defunct Cemetery
at the Church of St. John in Obora (Prague-Lesser Town) (in Czech).
Archeologie ve středních Čechách
, 16/2, 983–1019.
OMELKA, M., ŘEBOUNOVÁ, O., 2012b. A view of the archaeological
context of the Lesser Town cemetery in Šporkova Street in Prague
using Modern period iconography and written sources.
Studies in Post-
Medieval Archaeology
, 4, 233–250.
OMELKA, M., ŘEBOUNOVÁ, O., 2014. Barokní mariánské kulty na Malé
Straně v zrcadle pražských archeologických nálezů náboženských medailí
(s přihlédnutím k situaci v Čechách a na Moravě) – Baroque Maria’s
Cults in the Lesser Town in the Mirror of Prague Archaeological Finds
of Religious Medals (with Consideration to the Situation in Bohemia and
Moravia).
Pražský sborník historický
, 42, 243–268.
OMELKA, M., ŘEBOUNOVÁ, O., 2016. Zboží pro chudé a bohaté –
„originály“ a dobové „padělky“ náboženských medailek na příkladu
nálezů z hrobových kontextů – Products for the Poor and for the Rich:
“Originals” and “Fakes” of Religious Medals on the Example of Finds
from Grave Contexts (in Czech).
Archaeologia historica
, 41, 309–325.
OMELKA, M., ŘEBOUNOVÁ, O., 2017. Stav a perspektivy bádání
novověkého pohřebního ritu v Čechách – The State and Perspectives
of Research into Modern-Age Funeral Rites in Bohemia (in Czech).
Archaeologia historica
, 42, 117–133.
OMELKA, M., ŘEBOUNOVÁ, O., ŠLANCAROVÁ, V., 2009. Soubor
křížků ze zaniklého hřbitova při kostelu sv. Jana v Oboře (Praha1 –
Malá Strana). I. Obecné formy kříže – A Collection of Crosses from
the Defunct Cemetery at the Curch of St. John in Obora, Prague–Lesser
Town. I. General Forms of Crosses (in Czech).
Archeologie ve středních
Čechách
, 13(2), 1001–1083.
OMELKA, M., ŘEBOUNOVÁ, O., ŠLANCAROVÁ, V., 2010. Soubor
křížků ze zaniklého hřbitova při kostelu sv. Jana v Oboře (Praha1 – Malá
Strana). II. Speciální kříže – A Collection of Crosses from the Defunct
Cemetery at the Curch of St. John in Obora, Prague–Lesser Town. II.
Special Crosses (in Czech).
Archeologie ve středních Čechách
, 14(1),
423–476.
OMELKA, M., ŘEBOUNOVÁ, O., ŠLANCAROVÁ, V., 2011. Špendlík
– před hradbou a za hradbou – The Pin within and without the Town (in
Czech).
Archaeologia historica
, 36, 23–540.
PÄÄBO, S., 1989. Ancient DNA: Extraction, characterization, molecular
clonig, and enzymatic amplifcation.
Proceedings of the National
Academy of Sciences of the United States of America
, 86(6), 1939–1943.
PÄÄBO, S., POINAR, H., JAENICKE-DESPRES, V., HEBLER, J.,
ROHLAND, N., KUCH, M., KRAUSE, J., VIGILANT, L., HOFREITER,
M., 2004.
Genetic analyses from ancient DNA. Annual Reviews of
Genetics
, 38, 645–679.
PINHASI, R., FERNANDES, D., SIRAK, M., CONNELL, S., ALPASLAN-
ROODENBERG, S., GERRITSEN, F., MOISEYEV, V., GROMOV, A.,
RACZKY, P., ANDERS, A., PIETRUSEWSKY, M., ROLLEFSON, G.,
JOVANOVIC, M., TRINHHOANG, H., BAR-OZ, G., OXENHAM, M.,
MATSUMURA, H., HOFREITER, M., 2015. Optimal Ancient DNA
Yields from the Inner Ear Part of the Human Petrous Bone.
PLOS One
,
10(6), e0129102.
SIMÓN, M., JORDANA, X., ARMENTANO, N., SANTOS, C., DÍAZ, N.,
SOLÓRZANO, E., LÓPEZ, J.B. GONZÁLEZ-RUIZ, M., MALGOSA,
A., 2011. The presence of nuclear families in prehistoric collective burials
revisited: The bronze age burial of montanissell cave (spain) in the light
of aDNA.
American Journal of Physical Anthropology
, 146(3), 406–413.
TIERNEY, S., BIRD, J., 2014. Sex identifcation of human remains from
an Irish Medieval population using biomolecular methods.
European
Scientifc Journal
, 2, 521–530.
YANG, D.Y., ENG, B., WAYE, J.S., DUDAR, J.C., SAUNDERS, S.R.,
1998. Improved DNA extraction from ancient bones using silica-based
spin columns.
American Journal of Physical Anthropology
, 105(4), 539–
543.
YANG, D.Y. WATT, K., 2005. Contamination controls when preparing
archaeological remains for ancient DNA analysis.
Journal of
Archaeological Science
, 32(3), 331–336.
Other Sources
Prague City Archives, Collection of Registries, sign MIK Z3, MIK Z4,
MIK Z5.
Prague City Archives, Manuscript Collection, sign 4764.
The National Heritage Institute in Prague, Archive of the Department of
Archaeology, Documentation of excavations no. 30/02 and 30/04.