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TitleDNA Fingerprinting: State of the Science
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LanguageEnglish
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225

The micro satellite DNA markers used in this study are of the GAT A
type located on chromosome 12 (4804 LR and 4815 LR) and Y (27H39
LR) (Roewer et aI., 1992). 6 alleles are described for 4804LR, 8 for
4815LR and 4 for the 27H39LR locus in a number of caucasoids
(Roewer et aI., 1992; and Tab. 1). Because of the shortness of the PCR
amplified length variable alleles informative typing of old and degraded
DNA is possible. The sensitivity of PCR allows the collection of hairs
instead of blood for discriminative DNA analysis. HLA class II genes
are analyzed by the PCR-based oligotyping method.

Materials and methods

Genomic DNA was extracted from 3-10 hairs by an "in-one-tube
extraction" method (Kawasaki, 1990). Repeat flanking oligonucleotide
primers for the (GAT A)n micro satellite loci were used as described
previously (Roewer et aI., 1992). Approximately 100-250 ng DNA was
used for a 25-J.LI PCR sample with 2 U Taq polymerase in the buffer
recommended by the suppliers (Promega). The MgCl2 concentration
was 2.5 mM. Amplifications were performed for 30 cycles with 30 sec
denaturation at 94°C, 30 sec primer annealing at 51 °c and 90 sec
extension at 72°C in a PCR thermocycler (Biomed). Simultaneous
multilocus PCR using the 3 primer pairs was carried out under identical
conditions. Gel purification of the repeat-containing fragments, radio-
active labelling of the fragments, PAGE and X-ray exposure were
performed as previously described (Roewer et aI., 1991). A standard
ladder of the known alleles was used for sizing the bands.

HLA class II oligotyping (DQA 1 and DQB 1 loci) was done following
the protocols of the 11th International Histocompatibility Workshop in
Yokohama (1991).

Results

89 individuals were analyzed at the (GAT A)n micro satellite loci
4804LR, 4815LR (both linked on chromosome 12) and 27H39LR
(chromosome V). 75 individuals originate from the village HAP, the
remaining ones stem from the surrounding villages W A W, SHI, ARI,
MAR and MAA (Tab. 2).

In HAP 33 males and 42 females live in 3 generations. We found
several polygyneous families and 2 polyandreous families. 23 individuals
of the ancestral generation, who are possibly also more or less related,
were chosen for the calculation of observed allele frequencies, het-
erozygosity rates and gene diversities (Nei, 1973) (see Tab. 1). The

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226

Table 2. Microsatellite typing of 89 Yanomami indians from 6 villages (F father; M mother; C child;
m male; f female; 04 locus 4804LR; 15 locus 4815LR; Y locus 27H39LR); closely related individuals
are put to groups if possible

HAP Sex 04/15 /Y HAP Sex 04/15 /Y

1 F m ee/EF /a 44 F m ee/EF /a
2 M f cd/DF /- 45 M f be/ F/-
3 CI m cd/DE/a 46 CI f be/DF /-
4 C2 m cd/DE/a 47 C2 f ee/FF /-
5 C3 f cd/DF /- 48 C3 m ee/DF /a
6 C4 f cd/DE/- 49 C4 ee/FF /-

7 F m cc/DF /a 50 M f ee/EE /-
8 M f be/EF /- 51 CI f ee/EE /-
9 CI m ee/DF /a 52 C2 m ee/ /a

10 C2 m be/ED/a
53 m ee/FF /a

II F m ee/EE /a 54 m ee/FG /a
12 M f ee/EF /- 55 m be/EF fa.
13 CI f ee/EF /-
14 C2 m ee/EE /a 57 M f ee/FF /-
15 C3/M f ee/EG /- 58 Cl m ee/ /a
16 F m ee/FF /a 59 C2 f be/ /-
17 Cl/F m ee/FG/a 60 f ee/ /-
18 M f ee/EG/- 61 m cd/DF fa.
19 C f ee/GG/- 62 f be/ /-

63 F m ee/ /a
20 M f cd/EE /- 64 M f ee/ /-
21 Cl f ee/ /- 65 C m / /a
22 C2 f ee/EF /- 66 f ee/EG/-
23 C3 f cd/ /- 67 f cd/ /-
24 C4 m / /a 68 f ee/ /-
25 C5/M f cd/EF /- 69 f ee/EF /-
26 Cl m ee/ /a 70 m / /
27 C2 f ee/EF /- 71 f ee/ /-
28 C3 m cd/EF IP 72 m eel /a

73 f / /-
56 f cd/DF /- 74 f ee/ /-

75 f ee/ /-
29 F m eel D/a
30 M f ce/ /- WAW
31 Cl m ee/ / 1 m /a
32 C2 f ee/ 1- 4 m / /P
33 C3/M f ee/EF 1- 7 F m ee/EF /a
34 F m ee/ /a 8 M f ee/DE/-
35 Cl m ee/EF /a 9 Cl f ee/DF /-
36 C2 f ee/ /- 10 C2 f ee/DE/-
37 C3 m ee/ /a 13 m / fa.
38 C4 f ee/ /-
39 C5 f ee/ /- MAH
40 C6 m ee/EF / I m /P

4 m /a
41 F m be/CE /a II m /a
42 Cl f be/EG/- 19 m /a
43 C2 m ce/CG/a

MAA

m /P

ARI

m /a

SHI

m fa.

Page 452

B R K H A u 5 E R

L I F E SCIENCES
Experientia Supplementum

DNA Methylation:
Molecular Biology and
Biological Significance

Edited I¥

J.-P. Jost, Friedrich-Miescher Inst., Basel, Switzerland
H.-P. Saluz, IRBM, Rome, Italy

1993.572 pages. Hardcover. ISBN 3-7643-2778-2 (EXS 64)

•... This publication is timely and up-to-date and provides a comprehensive background not
available in other single sources . ... After reading it, I appreciate what a bargain that was .
... It is so recommended. • John C. Rogers, Washington Univ., st. louis, MO, USA

"Cell", May 7,1993

The goal of this book is to provide a comprehensive collection of reports on the state
of the art in DNA methylation. DNA methylation is found in many different genes
in bacteria, viruses, fungi, vertebrates and plants. It is involved in the protection of
DNA against restriction enzymes, X-chromosome inactivation, imprinting, changes
in DNA and chromatin structure, changes in the interaction of proteins with DNA,
Silencing viruses, and in embryogenesis and cancer.

This book gives students and newcomers to the field a unique introduction to the
topic and is also designed to help the experienced scientist to broaden and update
his knowledge in this highly topical field. The numerous references cited in the 24
chapters and the many explanatory figures and tables which complement the text
will guide and stimulate the investigator to further studies.

Please order through your bookseller or directly from:
Blrkhiuser Verlag AG, P.O. Box 133,
CH-4010 Basel 1 Switzerland (Fax +-+41/611 n1 7950)
Orders from the USA or Canada should be sent to:
'Blrkhiuser Boston
44 Hartz Way, Secaucus, NJ 07096-2491 1 USA
Call Toll-Free 1-800-V1-4643

Birkhiiuser i
BlrkhlllHr "'rIas AG
Buel . Boston . Berlin

Page 453

B R K H u 5 E R

L I F E SCIENCES
Advances in Life Sciences

Transgenic Organisms -
Risk Assessment of
Deliberate Release

Edited by

K. WOhnnann I J. Tomiuk. University of TObingen, Germany

1993. 280 pages. Hardcover. ISBN 3-7643-2834-7 (ALS)

The genetic structure of organisms is being altered to an ever-increasing degree by
molecular genetic techniques. Thus far, more than 500 transgenic organisms have
been tested or utilized in the field. The strides forward, however, have been
encumbered by persistent and intense controversy. What is happening now on the
forefront of genetic engineering? What are its aims, and inherent risks? What is its
justification?

The purpose of this book is to offer the reader a firm grounding in the subject by
systematically reviewing several basic issues: the interaction between host DNA!
RNA and the introduced DNAlRNA; the extent and effect of horizontal gene
transfer; the fate in populations of genes inadvertently released into nature; the
feasibility and effectiveness of risk assessment; and the manner in which this
modern technique can best be harnessed to improve human health. More generally,
but perhaps most significantly, the book calls for a critical appraisal of the desirability
of the aims advanced by molecular geneticists today.

Please order through your bookseller or directly from:
Blrkhiiuser Verlag AG, P.O. Box 133,
CH-4010 Basel I Switzerland (Fax ++41/611 n1 7950)
Orders from the USA or Canada should be sent to:
Blrkhiiuser Boston
44 Hartz Way, Secaucus, NJ 07096-24911 USA
Call Toll-Free 1-800-777-4643

Birkhiiuser i
Blrkhluser Verla. AG
B_1· Boston' Berlin

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