J Forensic Sci, 2016

doi: 10.1111/1556-4029.13246
TECHNICAL NOTE Available online at: onlinelibrary.wiley.com

CRIMINALISTICS

Sabine Hess,1 M.Sc.; and Cordula Haas,2 Ph.D.

Recovery of Trace DNA on Clothing: A

Comparison of Mini-tape Lifting and Three
Other Forensic Evidence Collection
Techniques

ABSTRACT: Trace DNA is often found in forensic science investigations. Experience has shown that it is difficult to retrieve a DNA profile
when trace DNA is collected from clothing. The aim of this study was to compare four different DNA collection techniques on six different
types of clothing in order to determine the best trace DNA recovery method. The classical stain recovery technique using a wet cotton swab
was tested against dry swabbing, scraping and a new method, referred to as the mini-tape lifting technique. Physical contact was simulated with
three different perpetrators on 18 machine-washed garments. DNA was collected with the four different DNA recovery methods and sub-
jected to standard PCR-based DNA profiling. The comparison of STR results showed best results for the mini-tape lifting and scraping methods
independent of the type of clothing. The new mini-tape lifting technique proved to be an easy and reliable DNA collection method for textiles.

KEYWORDS: forensic science, contact stains, trace DNA, touch DNA, DNA collection technique, mini-tapes, crime scene investigation,
DNA analysis

Trace or touch DNA refers to the DNA that is recovered from
skin (epithelial) cells that is left behind when a person touches
or comes into contact with items such as clothes, weapons, or
other objects. These cells are typically recovered when force is
used, such as on the victims clothes or at a crime scene, after a
struggle has occurred. The analysis of trace DNA opens new
dimensions in crime scene investigation. The availability of
DNA from touched objects assists in a wide range of offenses
including theft, armed robberies, volume crime, assaults, sexual
offenses, and homicides (1). A number of factors influence the
success rate of obtaining a DNA profile from the handled sub-
strate. In favorable conditions, handling time can be neglected,
as the transfer of DNA from sloughed epithelial cells to a sub-
strate during handling is instantaneous. Skin makes up 15% of
total body weight and is the largest organ of the human body.
The average human being sheds approximately 400,000 skin
cells a day (2). Individuals are thought to be either good or bad
shedders (2). Hands that have been recently washed provide little
DNA. Opinions differ on whether dry hands influence DNA
shedding (3,4). Offenses, especially crimes against the person,
often generate contact between skin and clothing. Experience
has shown that it is difficult to detect exploitable DNA profiles
when trace DNA is collected from clothing. Furthermore, the
surface of the handled textile influences the success rate for
1 recovery method is cutting out a suspected DNA-bearing section.
Forensic Science Institute Zurich, P.O. Box, 8021, Zurich, Switzerland.
2
Zurich Institute of Forensic Medicine, Forensic Genetics, University of
Zurich, Winterthurerstrasse 190, 8057, Zurich, Switzerland.
Received 1 Nov. 2015; and in revised form 7 Mar. 2016; accepted 24
April 2016.
obtaining a DNA profile. Sloughed epithelial cells adhere better
to porous substrates than to nonporous substrates (2). Because of
the irregular surface structure of natural fibers, it is expected that
epithelial cells will adhere better to those substrates than to syn-
thetic fibers. Furthermore, our experience has shown that it is
harder to retrieve exploitable DNA profiles from dark than from
bright textiles. This observation has been confirmed in a previ-
ous study where a lower amount of alleles was detected on dark
indigo cotton (jeans) than on white cotton (T-Shirt), which may
be due to the presence of indigo dye inhibiting PCR (5). This
was not only the case when DNA was extracted and amplified
from a cutout but also from a swabbed DNA-bearing fabric
section.
The success rate of obtaining a DNA profile from contact
traces depends mainly on the selection of the appropriate recov-
ery method for biological material and the way it is applied. The
first step in collecting trace DNA is to target the relevant area.
Even when using forensic light sources, trace DNA on clothing
is normally not visible. Therefore, trace DNA is mainly recov-
ered based on assumptions about where the DNA-containing
material could be located (1).
Many crime scene investigators and laboratories use wet
swabs to collect biological material, whereas deionized water is
mainly used as a moistening agent. It is also common practice to
perform a double-swabbing dry/wet technique, especially to
recover saliva on human skin (6). Another commonly used DNA
This technique is often used on soft items such as clothing
(1,7,8). Several DNA laboratories also use the scraping method
where the biological material is scraped off using a sterile
scalpel (8,9). A more recent method is the use of adhesive

2016 American Academy of Forensic Sciences 1

2 JOURNAL OF FORENSIC SCIENCES
mini-tapes for DNA recovery. In the past, this technique has
been commonly applied by forensic practitioners on the crime
scene as well as in the laboratory when collecting and preserving
trace evidence such as fibers and hairs. This recovery method is
quick, easy to apply, cost-effective, and ensures that invisible
traces are collected and well preserved (7,1012). A previous
study has demonstrated that the taping method is more suitable
for recovering cells from porous materials than swabs. However,
on nonporous materials, swabs showed a better performance than
the mini-tapes (11).
Materials and Methods
In this study, four different trace DNA collection methods on
six different types of clothing were tested. Trace DNA was
deposited on machine-washed used clothing by three volunteers
(standing for the perpetrators) on clothing from three owners
(representing the victims). For each perpetrator, a set of six
different types of clothing was provided. Trace DNA, which was
placed on a total of 18 garments, was then collected with the
four different techniques and subjected to DNA profiling.
Substrates and Contact Simulations
Six types of used clothing were utilized (three garments of
each type), namely blue jeans, raincoats, dark synthetic fabrics,
bright synthetic fabrics, dark colored natural fiber clothing, and
light colored natural fiber clothing (Table 1). Bright textiles were
white, beige, or light purple, whereas dark fabrics were black,
dark purple, or dark blue. Blue jeans, made from cotton which is
also a natural fiber type, were handled as a separate category as
they are very often encountered in casework. Raincoats were
selected as representatives of the nonporous textile surfaces
group. They were made of synthetic fibers with a PVC coating.
Reference profiles of the three owners, representing the vic-
tims (two males/V1 and V2; one female/V3), were available.
To remove as much preexisting biological material as possible,
all clothes were washed twice, together, in the same washing
machine. Each load of laundry was washed with 70 mL laundry
detergent, at 40C for 70 min, which is the most frequently used
washing program for delicate fabrics such as clothing. Similar
washing conditions were used by others (13,14). The laundry
was then air-dried and packed with appropriate care (gloves,
mask) to avoid contamination.
When simulating an assault, three individuals, not the owners
of the clothing, standing for the perpetrators (two males/P1
and P2; one female/P3) grabbed the clothing with both hands for
approximately 5 sec (Fig. 1). It was assumed that in a dynamic
process such as an assault, perpetrators can only hold onto the
same area on a garment of a resisting victim for a short period
of time. In order to be close to reality, the time period of touch-
ing the substrate was therefore chosen to be short. Between
applications, a time interval of at least 15 min was maintained,
TABLE 1The types of clothing used in the experiment, subdivided into substrate groups.
Substrate Subgroups Fabric Type
Bright natural fibers Cotton
Dark natural fibers Cotton
Bright synthetic fibers Acrylic, Nylon
Dark synthetic fibers Acrylic, Nylon, Viscose
Blue jeans Cotton
Raincoats Polyester, Nylon with PVC coating
FIG. 1Simulated contact of P1 () with piece of clothing. The clothing
was grabbed with both hands for approximately 5 sec. Between applications,
a time interval of at least 15 min was maintained.
without washing hands to enable best skin transfer conditions
(15). Except for the raincoatswhere the scraping method is not
routinely used and therefore only three methods were tested
the contact stains were deposited as triplicates in four places on
the five types of clothing to apply each sampling technique and
to check for reproducibility. This procedure was repeated three
times for each perpetrator which made a total of 207 deposits.
All the contact zones of approximately 3040 cm2 were circled
for DNA recovery.
On the 18 garmentsthree garments per substrate groupan
untouched area was analyzed as a blank. One Scenesafe mini-
tape was analyzed as a control sample.
Trace DNA Recovery Techniques
Four recovery methods were used to collect trace DNA from
the clothing, namely dry swabbing, wet swabbing, mini-tape lift-
ing, and scraping. When dry swabbing, a dry cotton swab was
rubbed with moderate pressure and rotation on the touched area
and stored in a cardboard box. The wet swabbing method was
similar in application, but a moistened (with sterile water) cotton
swab was used instead. The foldable cardboard boxes used in
this study have been especially designed for DNA recovery and
are suitable for drying and storage of epithelial cells on dry and
moistened cotton swabs. At room temperature, swabs completely
air dry within 69 h (16). Using the mini-tape lifting technique,
trace DNA was collected on the clothing using adhesive tapes
and these were transferred into a sterile 1.5 mL tube. When
applying the scraping technique, the surface of the touched area
of the substrate was scraped with a sterile scalpel blade and col-
lected material was transferred into a sterile 1.5 mL tube.
Color Garment Type Surface Structure
White T-Shirt, Sweatshirt Porous
Black, Dark blue T-Shirt, Sweatshirt Porous
Beige, Light purple Top, Jumper Porous
Black, Dark purple Cardigan, Jumper Porous
Dark blue Jeans Porous
Dark blue, Bordeaux Raincoat Nonporous

DNA-free ethylene oxide treated commercially available cot-
ton swabs (Prionics AG, Schlieren-Zurich, Switzerland) and Sce-
nesafe FASTTM mini-tapes (Scenesafe Ltd., Birmingham, U.K.)
were used.
DNA Analysis
Conventional Chelex extraction was performed according to
standard protocols (17,18), followed by ultrafiltration with Ami-
con Ultra-4 Centrifugal filter devices (Millipore, Zug, Switzer-
land) to a final extract volume of approximately 100 lL. 5 lL
of DNA was amplified with the SEfiler plus Multiplex Kit
(Applied Biosystems (AB), Rotkreuz, Switzerland) in a total
reaction volume of 25 lL on a GeneAmp PCR System 9700
(AB) according to the manufacturers protocol. The first samples
were additionally analyzed with the MiniFiler kit (AB), which is
beneficial for degraded DNA, but as good profiles were obtained
with the SEfiler plus kit, the MiniFiler data were not evaluated
further. PCR products were separated and detected with a 310
Genetic Analyzer (AB). Raw data were analyzed with the
Gene-mapper Software using a peak detection threshold of 50
RFUs. The numbers of detected alleles from each perpetrator
were compared to the numbers of maximal retrievable alleles per
perpetrator (percentage of detected alleles). The mean, stan-
dard deviation (SD), and relative standard deviation (%RSD)
were calculated for each evaluation, respectively. In addition, the
numbers of alleles that could be assigned to the victim or other
persons were evaluated.
Results
Perpetrators
Of the three perpetrators, P1 and P2 (males) were obviously
good shedders (90% and 82.6% of alleles were detected),
whereas P3 (female) was a poor shedder (only 14.2% of alleles
could be detected), irrespective of the clothing material, and
sampling method (Fig. 2). Perpetrators P1 and P2 (good shed-
ders, male) showed much lower %RSD than perpetrator P3 (poor
shedder, female) (Fig. 2).
FIG. 2Shedding capacity of the three perpetratorsafter contact with
different types of clothing, not considering the sampling method and the
clothing material. Twenty-three stains were analyzed in triplicates (n = 69)
per perpetrator (207 stains in total). The y-axis represents the percentage
of detected alleles from the respective perpetrator compared to the num-
ber of maximal retrievable alleles per perpetrator. Error bars represent
%RSD in relation to the mean of the DNA recovery rate of the correspond-
ing perpetrator. The numerical values of the RSD were 22%, 31%, and
126% for P1, P2, and P3, respectively.
HESS AND HAAS . RECOVERY OF TRACE DNA ON CLOTHING 3
Trace DNA Recovery Techniques
Four sampling methods were used to collect biological mate-
rial from clothing: dry swabbing, wet swabbing, mini-tape lift-
ing, and scraping. Comparison of STR results showed highest
DNA recovery for the mini-tape lifting and scraping sampling
methods, where 67% and 73% of alleles were detected, respec-
tively. Swabbing with dry or wet swabs provided slightly lower
recovery rates, namely 52% and 58%, respectively (data not
shown). The STR results of the perpetrators showed also high-
est DNA recovery for the mini-tape lifting and the scraping sam-
pling method, where 92% (P1/P2) or 18% (P3) and 93% (P1/P2)
or 32% (P3) of alleles were detected, respectively (Fig. 3).
Swabbing with dry or wet swabs provided slightly lower recov-
ery rates, namely 76% and 82% for P1 and P2, respectively. P3
showed rather lower DNA recovery rates, where 2% and 9% of
alleles were detected, respectively (Fig. 3).
The two DNA-sampling methods with higher DNA recovery
rates (scraping and mini-tape) showed generally lower relative
standard deviations (%RSD) than the two techniques with lower
DNA recovery rates (dry and wet swab) (Fig. 3).
For the raincoats, the dry swabbing and mini-tape methods
performed better (60% and 65% of alleles detected) than the wet
swabbing technique (51% of alleles detected). As the scraping
method is not routinely applied to nonporous surfaces, this
method was not used on the raincoats (Fig. 4).
In addition, the variation of triplicate analyses was calculated
for all simulated stains. The %RSD of the triplicates varied enor-
mously, from 0% to 173% (data not shown). This was observed
irrespective of the sampling method.
Clothing Material
Contact stains from dark natural materials (blue jeans
included) seemed to provide slightly better results than from syn-
thetic materials, irrespective of the sampling method and the
perpetrator. DNA results from bright natural material were
FIG. 3Performance of four DNA recovery techniques irrespective of the
substrate, grouped in good shedders (P1/P2) and poor shedder (P3). Eigh-
teen stains were collected in triplicates with dry swab, wet swab, and mini-
tape and 15 stains with scraping (207 stains in total). The y-axis represents
the percentage of detected alleles compared to the number of maximal
retrievable alleles. Error bars represent the relative standard deviation
(%RSD) in relation to the mean of the corresponding DNA recovery tech-
nique and perpetrators group. The numerical values of the RSD of the cor-
responding perpetrators shedding group (P1/P2 or P3) were 36% or 252%,
25% or 115%, 22% or 87%, and 20% or 76% for the dry swab, wet swab,
mini-tape, and scraping methods, respectively.
4 JOURNAL OF FORENSIC SCIENCES
FIG. 4DNA results after applying four different sampling methods on
four different substrates. Twenty-four stains were collected in triplicates
(n = 72) from synthetic and natural fiber textiles and 12 (n = 36) and 9
stains (n = 27) from blue jeans and raincoats, respectively (207 stains in
total). The y-axis represents the percentage of detected alleles compared to
the number of maximal retrievable alleles.
slightly worse than from bright synthetic material though. Good
shedders (group P1/2) performed better on bright synthetic than
on dark synthetic material, whereas bad shedders (group P3) pro-
vided better results on dark synthetic than on bright synthetic
material. On dark synthetic material, good shedders (group P1/2)
performed worse, whereas bad shedders (group P3) provided
worst results on raincoats (Fig. 5) irrespective of the sampling
method.
Other Findings
From a total of 207 analyzed stains, about 40% showed addi-
tional alleles, not belonging to the perpetrators (DNA mix-
tures). As all clothes were washed twice, STR profiles from the
victims were not expected. Nevertheless, six mixed STR pro-
files could be assigned to the perpetrator and the victim,
FIG. 5Performance of DNA recovery on six different substrates grouped
in good shedders (P1/P2) and poor shedder (P3), irrespective of the sam-
pling method. Twelve stains were collected in triplicates from five different
textile substrate groups (n = 36) and nine stains from raincoats (n = 27,
respectively, 207 stains in total). The y-axis represents the percentage of
detected alleles compared to the number of maximal retrievable alleles.
Error bars represent the relative standard deviation (%RSD) in relation to
the mean DNA recovery rate of the corresponding substrate and perpetra-
tors shedding group. The numerical values of the RSD of the corresponding
perpetrators group (P1/P2 or P3) were 20% or 110%, 51% or 147%, 29%
or 167%, 7% or 92%, 19% or 102%, 20% or 143% for the six different sub-
strates (bright synthetic fibers, dark synthetic fibers, bright natural fibers,
dark natural fibers, blue jeans, and raincoats, respectively).
meaning that more than four alleles uniquely belong to the vic-
tim. Several alleles and combination of alleles of unknown indi-
viduals appeared in addition to the perpetrators DNA profile,
some of them repeatedly. In two samples, one in the research
involved staff member was a contributor to the mixtures. One
sample was identified as single tube contamination (listed in the
DNA Elimination Database EDB) (19), and three minor contrib-
utors could not be assigned to known individuals. One STR pro-
file, which also could not be assigned to any known person,
appeared repeatedly in stains from two specific pieces of cloth-
ing. Of the 18 randomly selected blanks, two could be assigned
to the victim.
Discussion
The performance of four trace DNA collection techniques (dry
swabbing, wet swabbing, scraping, and mini-tape lifting) on six
different types of clothing (bright natural fibers, dark natural
fibers, bright synthetic fibers, dark synthetic fibers, blue jeans,
and raincoats) were compared in this study by means of exploi-
table STR profiling results.
Of the three perpetrators, the two males were good shed-
ders, whereas the female was a poor shedder. Previous studies
have demonstrated that there was no significant difference
between males and females regarding their shedding capacity
(20). Due to the small number of perpetrators in this study, no
conclusive reasoning is possible in this regard. Perpetrators P1
and P2 (good shedders, male) showed much lower %RSD than
perpetrator P3 (poor shedder, female). This can be explained by
the smaller mean number of detected alleles of P3, which makes
being the denominatorthe %RSD increase dramatically (21).
Therefore, the DNA recovery techniques were analyzed sepa-
rately for good and poor shedders.
The comparison of STR results showed the highest DNA
recovery rates for the mini-tape lifting and scraping sampling
methods, independent of the substrate. Low DNA recovery was
observed for the dry swabbing method, which demonstrates the
need of the moistening agent for dissolving epithelial cells from
a substrate. An exception was illustrated by the category of rain-
coats, the representative for nonporous surfaces, where dry
swabbing performed better than wet swabbing. A possible expla-
nation could be that dirt and inhibitors are easily collected and
dissolved in water, which would be a drawback of the wet swab-
bing method. Nevertheless, the mini-tape method also showed
the best DNA recovery for raincoats.
Results from triplicate analyses showed that DNA recovery
was highly variable (%RSD up to 173%), irrespective of the
sampling method. As the sampling and analysis methods were
performed in a reproducible way, the reason for this outcome is
most probably the difficulty of placing identical stains on cloth-
ing by grabbing, a method which is influenced by many noncon-
trollable factors (such as pressure and amount of DNA cells
present on hands of perpetrator before simulated contact).
Overall, trace DNA collected from natural dark fiber materials
(including blue jeans) revealed slightly more complete DNA pro-
files than from synthetic materials (including raincoats). There-
fore, sloughed epithelial cells might adhere better to porous
surfaces such as natural fiber materials than to both nonporous
surfaces and synthetic fiber materials. This would be consistent
with a previous report, which demonstrated that DNA cells
adhere far better to swabs made from cotton rather than synthetic
fibers (2). The fact that the DNA recovery rate on bright natural
materials was slightly worse than on bright synthetic materials is

not consistent with this theory though. As there was only one
garment per type of substrate and perpetrator used in this
study, no conclusive reasoning is possible in this regard.
Concerning the color of the textiles, good shedders (P1/2) per-
formed better on bright synthetic fiber material than on dark syn-
thetic fiber material, whereas bad shedders (P3) provided better
results on dark synthetic than on bright synthetic fiber material.
Overall good and bad shedders performed better on dark natural
fiber material (including blue jeans) than on bright fabrics. We
conclude that the color of the clothing did not have an impact
on the DNA recovery rate in this study, which is neither consis-
tent with our experience in casework nor with previous research
(5). This could be due to the four DNA recovery methods tested
(wet swabbing, dry swabbing, scraping, and tape lifting), where
only little colored fiber material is collected together with the
epithelial cells. However, when DNA is extracted from a cutout
piece of fabric, there is more dyed fiber material or rather dye
present which could inhibit the PCR (5,22). Alternatively, this
observation could be a result of small sample size.
On all six types of clothing, mixed DNA profiles were
retrieved, namely the perpetrator and an additional person. In
spite of washing the clothes twice before trace DNA deposition,
in six of 207 stains, there was still residual DNA of the victim
found. Not all involved persons in the mixtures could be identi-
fied. For example, one DNA profile was observed repeatedly on
two pieces of clothing from the same victim. This reflects real-
ity and shows the need for collecting DNA from all involved
persons (people living in the same household, police staff, prose-
cutors, medical staff, etc.) who could have legitimately left their
DNA on evidence items.
Conclusion
More complete DNA profiles were retrieved when either the
mini-tape lifting or the scraping method was used to collect trace
DNA from clothing. However, mini-tapes are easier and faster to
apply when minute biological trace evidence is recovered at
crime scenes and in the laboratory. The mini-tape lifting method
is now routinely used at the Forensic Science Institute Zurich to
collect trace DNA on textile substrates such as clothing, car
seats, and other fabrics. An improvement in the success rate of
obtaining exploitable DNA profiles from trace DNA on textile
surfaces has been observed when applying this method in case-
work. Whereas previously full DNA profiles were hardly ever
retrievable from contact traces on garments, full and exploitable
DNA profiles from perpetrators having touched clothing and
other textile surfaces are now routinely possible.
Acknowledgments
Anna Jelena Lengacher is acknowledged for carrying out part
of this project during her internship in Zurich. The authors
would also like to thank Marion Walt for all the laboratory
work. The authors would like to extend the thanks to all the col-
laborators of the Forensic Science Institute Zurich and the Insti-
tute of Forensic Medicine who have contributed to this study.
Finally, the critical review of the final version and linguistic
revision by Dr. Michael Taylor from the New Zealand Institute
HESS AND HAAS . RECOVERY OF TRACE DNA ON CLOTHING 5
of Environmental Science and Research are very much
appreciated.
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Additional information and reprint requests:
Sabine Hess, M.Sc.
Forensic Science Institute Zurich
P.O. Box
8021 Zurich
Switzerland
E-mail: sabine.hess@for-zh.ch