Forensic Science International: Genetics 16 (2015) 8–12

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Forensic Science International: Genetics

journal homepage: www.elsevier.com/locate/fsig

Short Communication

ESDA1-Lite collection of DNA from latent fingerprints on documents

Dane T. Plaza *, Jamia L. Mealy, J. Nicholas Lane, M. Neal Parsons, Abigail S. Bathrick,
Donia P. Slack
The Bode Technology Group, 10430 Furnace Road, Suite 107, Lorton, VA 22079, USA

A R T I C L E I N F O A B S T R A C T

Article history: The ability to detect and non-destructively collect biological samples for DNA processing would benefit
Received 22 July 2014 the forensic community by preserving the physical integrity of evidentiary items for more thorough
Received in revised form 8 October 2014 evaluations by other forensic disciplines. The Electrostatic Detection Apparatus (ESDA1) was
Accepted 9 November 2014
systemically evaluated for its ability to non-destructively collect DNA from latent fingerprints deposited
on various paper substrates for short tandem repeat (STR) DNA profiling. Fingerprints were deposited on
Keywords: a variety of paper substrates that included resume paper, cotton paper, magazine paper, currency, copy
DNA typing
paper, and newspaper. Three DNA collection techniques were performed: ESDA collection, dry swabbing,
ESDA
Non-destructive
and substrate cutting. Efficacy of each collection technique was evaluated by the quantity of DNA present
Evidence collection in each sample and the percent profile generated by each sample. Both the ESDA and dry swabbing non-
Latent fingerprints destructive sampling techniques outperformed the destructive methodology of substrate cutting. A
DOMEX greater number of full profiles were generated from samples collected with the non-destructive dry
swabbing collection technique than were generated from samples collected with the ESDA; however, the
ESDA also allowed the user to visualize the area of interest while non-destructively collecting the
biological material. The ability to visualize the biological material made sampling straightforward and
eliminated the need for numerous, random swabbings/cuttings. Based on these results, the evaluated
non-destructive ESDA collection technique has great potential for real-world forensic implementation.
ß 2014 Elsevier Ireland Ltd. All rights reserved.

1. Introduction
The ability to successfully detect, collect, and process individual
biological samples from various evidentiary substrates without
causing surface damage is a challenge in the field of forensics. The
damage inflicted to an evidence item during sample collection may
prevent additional evaluations of the object. Traditional methods
of recovering DNA from forensic samples typically rely on the use
of chemical sprays, wet/dry cotton tip swabbing, or substrate
cuttings [1–7]. While these traditional techniques effectively
collect biological material, they can leave evidence items in an
altered or damaged state. Chemical sprays, such as Ninhydrin, are
utilized frequently by forensic investigators in order to locate
amines left behind by sloughed off cellular debris [8,9], but the use
of this chemical may destroy the integrity of the evidence,
rendering it unusable for further forensic testing. The wet/dry
double swab technique is a highly employed collection method
used for the sampling of biological deposits on a wide variety of
* Corresponding author. Tel.: +1 703 646 9740; fax: +1 703 646 9741.
E-mail address: dane.plaza@bodetech.com (D.T. Plaza).
items, but this methodology typically causes tearing and blotting
when used to sample paper documents. Substrate cuttings provide
DNA analysts with small clippings of the original sample that can
yield ample amounts of DNA through the extraction process [6];
however, this type of sampling is also very destructive as it
involves physically removing segments of evidence. Although
these sampling techniques represent effective means of collecting
DNA, they are often destructive and can damage evidentiary items,
which limits their usefulness to other forensic disciplines.
Non-destructive collection techniques are needed in the forensic
community to supplement established destructive methods of
sampling biological material. Forensic DNA analysts would benefit
from a technique that allows for the detection and collection of
biological materials from an item without damaging its structural
integrity or interfering with any subsequent examinations. The
additional information gained from questioned documents, entry
point surfaces, various clothing items, and other items containing
touch DNA evidence processed in this manner could aid in the
conviction or exoneration of individuals. Evidence processed in a
non-destructive manner would also remain available for future
evaluations, which could prove pivotal to the outcome of a cold case
investigation or criminal retrial.

http://dx.doi.org/10.1016/j.fsigen.2014.11.011
1872-4973/ß 2014 Elsevier Ireland Ltd. All rights reserved.
 D.T. Plaza et al. / Forensic Science International: Genetics 16 (2015) 8–12
Ideally, the development of a non-destructive collection
technique will entail the use of instrumentation and/or methods
that are already accepted and used within the forensic community.
The Electrostatic Detection Apparatus (ESDA1) is a widely used
tool for forensic document examinations. This instrument was
developed for the recovery of indented impressions that are
created when writing occurs on a sheet of paper that is resting
upon other pages. The device applies an electrostatic charge over a
thin polymer, termed Mylar film, which is secured to an
evidentiary document by gentle vacuum suction [10]. The Mylar
film adheres to the form of the original document and highlights
discrepancies in the document surface, such as writing impres-
sions and latent fingerprints [10–13]. These markings become
visible to the naked eye when a developer, which consists of small
toner-covered glass beads that are attracted to the electrostatic
charge, is applied to the Mylar film that is then analyzed for
handwriting styles, replica text, or fingerprint markings while the
original document remains unscathed [10].
Because the Mylar film comes into contact with the original
document during the ESDA process, DNA has the potential to be
transferred from the evidence item to the film. Not only could
electrostatic detection techniques be used to collect DNA
evidence from documents that need to be preserved for further
analysis, but the ESDA may also be beneficial for collecting DNA
from latent fingerprints deposited on evidentiary items such as
documents. Traditionally, if fingerprints are not visible on an
item, multiple non-destructive dry swab samplings or destruc-
tive techniques are employed, whereas with the ESDA, finger-
prints, if present, would be visualized, making sampling
straightforward and efficient for targeted sampling of only the
areas of interest. The benefits of a non-destructive collection
technique warrant an evaluation of the ability of the ESDA to
obtain usable short tandem repeat (STR) profiles from documents
of interest. In this study, collections of latent fingerprints from
paper substrates of varying weight were compared between the
ESDA, dry swabbing, and destructive (i.e. substrate cutting and
wet/dry swabbing) collection techniques.
2. Materials and methods
2.1. Sample preparation
Fingerprints from three donors were deposited in triplicate on
paper substrates of varying weight: mid-weight resume paper,
cotton paper, magazine paper, and currency; standard weight copy
paper; and low-weight newspaper. All substrates were deconta-
minated by UV cross-linking prior to use. Donors were asked to
‘‘charge’’ their fingerprints by rubbing their fingertips over their
faces and necks to ensure that an abundance of epithelial cells and
sebaceous oils would be present in the deposited fingerprints. A
total of 162 samples were dried overnight at room temperature.
For each collection technique, 54 latent fingerprints were sampled.
2.2. Sample collection
Fifty-four fingerprints were collected with the ESDA collection
technique, which was performed utilizing the ESDA-Lite (Foster
and Freeman, United Kingdom), followed by wet/dry swabbing of
the portion of the Mylar film that came into contact with the
fingerprint. To prevent sample cross-contamination, samples were
not permitted to make direct contact with the vacuum bed of the
instrument. Before each sample was placed on the ESDA-Lite, a
clean sheet of copy paper was placed directly on the vacuum bed
(Fig. 1a), and the sample was placed on the paper. After this, the
vacuum was started, and Mylar film was carefully placed over the
sample (Fig. 1b). To initiate an electrostatic charge, the corona
9
wand was turned on and slowly waved horizontally and vertically
over the plate at a height of approximately three to five
centimeters (Fig. 1c). Once the Mylar film was charged, cascade
developer was poured over it, and the Mylar film was fixed with a
more rigid transparent fixing film so that the thin Mylar would not
fold over onto itself upon removal (Fig. 1d). Sample areas of
interest were marked for ease of collection, the vacuum was turned
off, and the sample was removed (Fig. 1e). To maximize the DNA
recovered from the Mylar film, wet/dry swabbing with a
HydraFlock1 sterile flocked swab (Puritan, Guilford, ME) was
used to sample the side of the Mylar that came into contact with
the biological material (Fig. 1f).
Dry swabbing was employed as an additional non-destructive
sampling technique to be compared to collection with the ESDA-
Lite. Fifty-four fingerprints were collected from the paper
substrates with Hydraflock swabs via direct dry swabbing of the
area where the fingerprint was applied. Finally, to allow for a
comparison of the non-destructive collection techniques to a
destructive collection technique, approximately one inch squared
cuttings of the paper substrates were taken from the areas where
the remaining 54 fingerprints were applied. Rather than deface the
currency by direct cutting of the substrate, the wet/dry swabbing
technique with HydraFlock swabs was utilized to sample the areas
where the fingerprints were deposited.
2.3. Sample processing
All samples were extracted using the Qiagen1 EZ11 DNA
Investigator Kit on the EZ1 Advanced Instrument (Qiagen, Venlo,
Limburg, The Netherlands). The wet/dry swabbed ESDA samples
were lysed separately, and the lysates were combined into a single
tube for extraction on the EZ1 instrument. Quantification was
performed with the Quantifiler1 Duo DNA Quantification Kit (Life
TechnologiesTM, Carlsbad, CA). Samples that contained less that
0.1 nanogram per microliter (ng/ml) of DNA were concentrated
with Vivacon1 500–30 K columns (Vivaproducts, Littleton, MA).
Amplification was performed with the AmpF‘STR1 Identifiler1
Plus PCR Amplification Kit (Life Technologies, Carlsbad, CA). A
template DNA concentration of 1 ng was targeted for each
amplification reaction (28 cycles, 12.5 ml reaction volumes).
Capillary electrophoresis was performed on the 3130xL Genetic
Analyzer (Life Technologies, Carlsbad, CA), and results were
analyzed using ABI GeneMapper1 v3.2.1 software (Life Technolo-
gies, Carlsbad, CA) with an analytical threshold of 50 relative
fluorescent units (RFU). Appropriate substrate negative controls,
extraction positives, reagent blanks, positive amplification con-
trols, and negative amplification controls were employed. JMP1
Software: Classic Design of Experiments (DOE) (SAS, Cary, NC) was
used to perform trend analysis of the data based on the effects of
multiple factors (i.e. substrate, collection method, and donor).
3. Results
The efficacy of each collection technique was evaluated by the
quantity of DNA present in each sample and the percent STR profile
(alleles obtained/alleles expected  100) generated by each
sample. For each collection technique and substrate type, the
DNA quantities were averaged across all three donors. Large
standard deviations were observed due to the wide range of DNA
quantities deposited by the different donors. The copy paper
samples were the only samples collected with the ESDA that
demonstrated a statistically significant difference (p < 0.05) in
DNA quantity when compared to those collected with dry
swabbing (Table 1). No other samples collected with the ESDA
technique demonstrated statistically significant differences in
DNA quantity when compared to the samples collected via dry
[(Fig._1)TD$IG]
10 D.T. Plaza et al. / Forensic Science International: Genetics 16 (2015) 8–12

Fig. 1. Samples were collected using (a) the ESDA-Lite. (b) The vacuum was started, and the Mylar film was placed over the sample, after which (c) the corona wand was used
to initiate an electrostatic charge. Once charged, (d) cascade developer was poured over the Mylar film, and the Mylar film was fixed with a transparent fixing film for easy
removal of samples. (e) The vacuum was turned off, and the sample was removed. (f) The bottom side of the Mylar containing the localized areas of biological material was
then wet/dry swabbed with a HydraFlock swab and processed for DNA.

swabbing. When compared to the fingerprint samples collected
with the destructive substrate cutting technique, the ESDA
samples collected from cotton paper demonstrated a statistically
significant increase in DNA quantity; however, when collecting
samples from newspaper, the ESDA collection technique resulted
in a statistically significant decrease in DNA quantity when
compared to substrate cutting. The samples collected with the dry
swabbing technique from resume paper, cotton paper, and
currency displayed statistically significant increases in DNA
quantity when compared to those collected with the destructive
collection techniques; however, when collecting samples from
newspaper, the dry swabbing collection technique resulted in a
statistically significant decrease in DNA quantity when compared
to substrate cutting.
When examining the percent profile generated by each sample,
the number of full and high partial profiles generated by each
collection technique was determined. Profiles in which 70% of the
alleles (22 of 32 alleles or 11 loci) were obtained were considered
to be high partial profiles. Across all substrates tested, full to high
partial profiles were obtained for 65% of samples utilizing the
non-destructive ESDA collection technique, 93% of samples
utilizing the non-destructive dry swabbing technique, and 52%
of samples collected using a destructive collection technique
(Table 2). Overall, non-destructive dry swabbing outperformed the
other two collection techniques for all donors on all types of
paper substrates except newspaper on which direct substrate
cutting worked best. Non-destructive collection with the ESDA
outperformed destructive substrate cutting on resume paper,

Table 1
Two-sample t-test comparisons of DNA quantity (ng) present in fingerprint samples collected via ESDA, dry swabbing, and destructive (material cutting or wet/dry swabbing)
collection techniques.

Paper substrate ESDA Dry swabbing Destructive p-value

X̄ SD X̄ SD X̄ SD ESDA vs dry ESDA vs Dry swabbing vs

swabbing destructive destructive

Resume 0.367 0.601 0.521 0.348 0.061 0.097 0.516 0.170 0.004*
Cotton 0.217 0.264 0.547 0.519 0.000 0.000 0.115 0.039* 0.013*
Magazine 0.332 0.308 1.049 1.186 0.130 0.149 0.113 0.102 0.050
Currency 0.320 0.696 0.623 0.319 0.187 0.184 0.260 0.592 0.003*
Copy 0.170 0.185 0.798 0.460 0.550 0.881 0.003* 0.237 0.468
Newspaper 0.170 0.226 0.267 0.245 0.677 0.453 0.400 0.011* 0.030*

Notes: *p < 0.05 (two-tailed test); X̄: mean; SD: standard deviation.
 D.T. Plaza et al. / Forensic Science International: Genetics 16 (2015) 8–12 11

Table 2
Full profiles generated from the fingerprint samples deposited on various paper substrates and collected via the ESDA-Lite, dry swabbing, and destructive (material cutting or
wet/dry swabbing) collection techniques.

Substrate ESDA collection Dry swabbing collection Destructive collection

N Full High partial Full and high N Full High partial Full and high N Full High partial Full and high
profiles profiles partial (%) Profiles profiles partial (%) profiles profiles partial (%)

Resume paper 9 3 3 67% 9 8 1 100% 9 0 1 11%

Cotton paper 9 4 3 78% 9 8 0 89% 9 0 0 0%
Magazine paper 9 3 4 78% 9 7 2 100% 9 2 4 67%
Currency 9 3 2 56% 9 8 1 100% 9 1 3 44%
Copy paper 9 5 2 78% 9 9 0 100% 9 5 3 89%
Newspaper 9 3 0 33% 9 5 1 67% 9 8 1 100%
Mid weight 36 13 12 69% 36 31 4 97% 36 3 8 31%
Standard weight 9 5 2 78% 9 9 0 100% 9 5 3 89%
Low weight 9 3 0 33% 9 5 1 67% 9 8 1 100%

Total 54 21 14 65% 54 45 5 93% 54 16 12 52%

Note: N: total samples.

cotton paper, and magazine paper. For both the non-destructive
ESDA and destructive collection techniques, results of collection
from currency differed slightly depending on the donor, with fewer
full and high partial profiles obtained from samples deposited by
donor 3.
4. Discussion
The ESDA collection technique successfully recovered sufficient
DNA from latent fingerprints on paper substrates to yield useable
STR profiles; however, large standard deviations were observed
when the DNA quantity results were averaged across all three
donors. This may be more indicative of the nature of fingerprint
deposition than the reliability of the collection techniques.
Replication of DNA results from fingerprint samples can be
challenging as the transfer and collection of DNA from fingerprints
is influenced by a number of factors, including donor perspiration,
shedder status, frequency of hand washing, tendency of the donor
to touch other areas of the body, pressure and friction during
contact, and type of substrate [4,14–16]. Of these factors, only
donor perspiration and shedder status cannot be mitigated though
instructions to the donor [4,15,16]. While the amount of DNA
transferred from a fingerprint to a substrate can be increased by
perspiration introducing additional epithelial cells and cell-free
nucleic acids to the surface of the skin [15,17], the amount of
epithelial cells an individual sheds has the largest impact on the
reproducibility of DNA results from fingerprint samples. Typically,
individuals can be categorized as high shedders, who shed more
epithelial cells, and low shedders, who shed fewer epithelial cells,
but it has been shown that fingerprints deposited by the same
donor on different days may demonstrate variability in the amount
of DNA that can be recovered [4,14,18].
Although some variation in DNA recovery was observed
between donors, it was shown that full or high partial DNA
profiles can be obtained from latent fingerprints by swabbing the
Mylar film after ESDA processing. In general, the dry swabbing
technique yielded the highest percentage of full and high partial
profiles. One factor that may have contributed to the success of the
dry swabbing technique is that it entailed a single transfer of
biological material from the document to the swab, whereas the
ESDA technique involved multiple biological material transfers:
one from the document to the Mylar, and a second from the Mylar
to the swab(s). The additional transfer step likely resulted in
decreased DNA recovery because some biological material was left
behind after each transfer. Additionally, this was a controlled
experiment in which latent fingerprints were deposited on known
locations on the substrates. Because dry swabbing was performed
on small areas that were known to contain biological material, the
likelihood of successfully collecting DNA was increased. In a real-
world scenario, the location of latent fingerprints on a document
may be unknown, resulting in decreased collection efficiency. In
this instance, the ESDA technique’s ability to visualize and collect a
sample may present a distinct advantage over dry swabbing.
Fingerprints have been successfully visualized with the ESDA up to
one month after being deposited on copy paper, glass, and 100%
cotton [D. Plaza, unpublished observation]. Furthermore, after
fingerprint visualization, additional DNA can be collected directly
from the document by noting the area where a fingerprint is
present and performing targeted direct dry swabbing of the
document’s surface. Although the ESDA collection technique was
outperformed by dry swabbing, it generated a higher percentage of
full and high partial profiles than substrate cutting across most
substrates. The ESDA collection technique was more effective
when collecting cellular material from latent fingerprints deposit-
ed on mid-weight paper substrates (i.e. cotton paper and magazine
paper), while direct cutting is better suited for collection of
fingerprints from low-weight paper substrates, such as newspaper.
Because DNA evidence is transferred to the Mylar film during
ESDA processing, practitioners may want to evaluate the sequence
in which a document will be examined by multiple disciplines.
During routine analysis, the ESDA is used to detect/visualize latent
handwriting indentations on paper documents, fixing film is
placed over the Mylar to create a lift that can be retained for further
evaluation, and then fingerprint treatment and DNA analysis are
performed on the original document. Alternatively, DNA evidence
can be collected from the Mylar immediately after the fixed image
is removed from the instrument. STR profiles can then be obtained
from the item in question while the structural integrity of the
evidence is preserved for further analyses. This creates a
streamlined process that maximizes the information obtained
from a single piece of evidence without interrupting the workflow
of the document or fingerprint examiners. Collection of DNA in this
manner would be particularly beneficial for historical documents,
currency, and documents that the prosecutor or submitting agency
requests to keep intact for trial. This technique was successfully
used to develop an investigative lead from a document that had
been handled by a person of interest less than a week prior to
processing. In this instance, the submitting party specifically
requested that the document remain unaltered. One full profile
that matched the reference profile was generated from a sample
collected with the ESDA technique.
5. Conclusions
The ability to visualize and non-destructively collect biological
samples will allow other forensic disciplines to perform thorough
12 D.T. Plaza et al. / Forensic Science International: Genetics 16 (2015) 8–12
forensic investigations of evidence items. Vital DNA evidence may
be lost when fingerprint, trace evidence, and chemical examina-
tions are performed in advance of biological inspections. Allowing
DNA analysts access to unprocessed evidence could increase the
likelihood of non-destructively collecting sufficient biological
material to produce a high quality DNA profile, particularly when
examining touch evidence items. For example, the use of an ESDA
to collect biological deposits from a handwritten document would
allow biological material to be recovered during the initial
processing of the item, and it would not interfere with subsequent
fingerprint development and/or handwriting analysis. The imple-
mentation of this technique would allow forensic scientists to gain
more information from sensitive evidence items by allowing
multiple full-scale examinations to be performed.
Typical document and media exploitation (DOMEX) techniques
for latent fingerprint detection and DNA collection can damage or
otherwise compromise the structural integrity of a document.
Alternatively, non-destructive collection can be performed by
sampling various sections of a document with numerous,
independent dry swabbings. Although the dry swabbing technique
can effectively recover DNA, it is used to sample random areas
where latent fingerprints are speculated to be deposited. ESDA
collection offers a unique advantage over dry swabbing in that it
allows for visualization of areas of biological relevance (i.e. latent
fingerprints) while facilitating non-destructive DNA collection.
As an alternative to destructive (e.g. chemical sprays, wet/dry
swabbing, substrate cutting) or standard non-destructive techni-
ques (e.g. dry swabbing), the ESDA is an effective tool for collection
of biological evidence from documents. Because DNA collection
takes place on the Mylar film, no destructive direct samples are
taken from the document, which is left completely intact. Full STR
profiles can then be generated from the samples. Additional
research on the ESDA’s ability to aid in non-destructive processing
of DNA from aged latent fingerprints on documents could be
extremely valuable for intelligence, law enforcement, and crime
laboratories.
Acknowledgements
This project was supported by Award No. 2010-DN-BX-K191,
awarded by the National Institute of Justice, Office of Justice
Programs, U.S. Department of Justice. The opinions, findings, and
conclusions or recommendations expressed in this publication are
those of the authors and do not necessarily reflect those of the
Department of Justice.
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