Abstract

Capillary electrophoresis (CE) is a versatile technology with tremendous potential applications in forensic science. A refinement of the traditional slab-gel, separation occurs in a liquid polymer contained within a capillary. The capillary electrophoresis unit is computer-driven, semi-automating the loading and analysis of samples.

As with any new technology, a full performance assessment is required before it may be implemented with confidence. This paper focuses on validation of CE technology for analysis of a portion of the X-Y homologous amelogenin gene used in gender determination. Typical Y/X ratios of peak heights for male samples were determined. Instrument linearity, sample resolution and reproducibility were examined. Samples subjected to contamination, extreme environmental conditions or extracted from a variety of substrates were also tested by CE. All samples typed correctly. Genetic material from a number of common non-primate animals was amplified with amelogenin primers. Some species yielded no product. Products derived from the animal samples that did amplify produced peaks on CE analysis readily distinguishable from those of human origin.

References

1.
Northrop
D
,
McCord
B
,
Butler
J
.
Forensic applications of capillary electrophoresis
.
J Capillary Electrophoresis
1994
;
1
:
158
-
68
.
2.
Butler
J
,
McCord
B
,
Jung
J
,
Lee
J
,
Budlowe
B
,
Allen
R
.
Application of dual internal standards for precise sizing of polymerase chain reaction products using capillary electrophoresis
.
Electrophoresis
 0173-0835
1995
;
16
:
974
-
80
.
3.
Buel
E
,
Schwartz
M
,
LaFountain
M
.
Capillary electrophoresis STR analysis: comparison to gel-based systems
.
J Forensic Sci
1998
;
43
(
1
):
164
-
70
.
4.
Nakahori
Y
,
Takenka
O
,
Nakagome
Y
.
A human X-Y homologous region encodes ‘Amelogenin.’
Genomics
 0888-7543
1991
;
9
:
264
-
9
.
5.
Bailey
D
,
Affara
N
,
Ferguson-Smith
M
.
The X-Y homologous gene amelogenin maps to the short arms of both the X and Y chromosomes and is highly conserved in primates
.
Genomics
 0888-7543
1992
;
14
:
203
-
5
.
6.
Buel
E
,
Wang
G
,
Schwartz
M
.
PCR amplification of animal DNA with human X-Y primers used in gender determination
.
J Forensic Sci
1995
;
40
:
641
-
4
.
7.
Gibson
C
,
Golub
E
,
Abrams
W
,
Shen
G
,
Ding
W
,
Rosenbloom
J
.
Bovine amelogenin message heterogeneity: alternative splicing and Y-chromosome gene transcription
.
Biochemistry
 0006-2960
1992
;
31
:
8384
-
8
.
8.
Lyngstadaas
S
,
Risnes
S
,
Nordbo
H
,
Flones
A
.
Amelogenin gene similarity in vertebrates: DNA sequences encoding amelogenin seem to be conserved during evolution
.
J Comp Physiol Bio
1990
;
160
:
469
-
72
.
9.
Watson
J
,
Spencer
J
,
Marshall Graves
J
,
Snead
M
,
Lau
E
.
Autosomal localization of the amelogenin gene in monotremes and marsupials: Implications for mammalian sex chromosome evolution
.
Genomics
 0888-7543
1992
;
14
:
785
-
9
.
10.
Akane
A
,
Seki
S
,
Shioni
H
,
Nakamura
H
,
Hasegawa
M
,
Kagawa
M
, et al
Sex determination of forensic samples by dual PCR amplification of an X-Y homologous gene
.
Forensic Sci Int
1992
;
52
:
143
-
8
.
11.
Akane
A
,
Shiono
H
,
Matsubara
K
,
Nakahori
Y
,
Seki
S
,
Nagafuchi
S
, et al
Sex identification of forensic specimens by polymerase chain reaction (PCR): two alternative methods
.
Forensic Sci Int
1991
;
49
:
81
-
8
.
12.
Casarino
L
,
De Stefano
F
,
Mannucci
A
,
Canale
A
.
HLA-DQA1 and amelogenin coamplification: a handy tool for identification
.
J Forensic Sci
1995
;
40
(
3
):
456
-
8
.
13.
Mannucci
A
,
Sullivan
K
,
Ivanov
P
,
Gill
P
.
Forensic application of a rapid and quantitative DNA sex test by amplification of the X-Y homologous gene amelogenin
.
Int J Legal Med
1994
;
106
:
190
-
3
.
14.
Roy
R
,
Steffens
D
.
Infrared fluorescent detection of PCR amplified gender identifying alleles
.
J Forensic Sci
1997
;
42
(
3
):
452
-
60
.
15.
Nakahori
Y
,
Hanamo
K
,
Iwaya
M
,
Nakagome
Y
.
Sex identification by polymerase chain reaction using X-Y homologous primers
.
Am J Med Gen
1991
;
39
:
472
-
3
.
16.
Sullivan
K
,
Mannucci
A
,
Kimpton
C
,
Gill
P
.
A rapid and quantitative DNA sex test: fluorescence-based PCR analysis of X-Y homologous gene amelogenin
.
Biotechniques
1993
;
15
:
636
-
41
.
17.
Buel
E
,
Schwartz
M
.
DAPI, a simple alternative to ethidium bromide staining of DNA in agarose gels
.
Appl Theor Electrophoresis
1993
;
3
:
253
-
5
.
18.
Luckey
J
,
Norris
T
,
Smith
L
.
Analysis of resolution in DNA sequencing by capillary gel electrophoresis
.
J Phys Chem
1993
;
97
:
3067
-
75
.
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