Forensic Science International 295 (2019) 145–149

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Forensic Science International

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

Technical Note

DNA recovery from gelatin fingerprint lifters by direct proteolytic

digestion
Martin Zieger* , Christoph Schneider, Silvia Utz
Institute of Forensic Medicine, Forensic Molecular Biology Dpt., University of Bern, Sulgenauweg 40, 3007 Bern, Switzerland

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

Article history: Fingerprints are a valuable source for DNA profiling in forensic investigations. In practice, the fingerprints are
Received 12 June 2018 routinely visualized first by powder staining and then often transferred to tapes or gelatin lifters for storage
Received in revised form 9 November 2018 or examination. If at all, fingerprints are usually sampled for DNA in a second step. To target the DNA sampling
Accepted 7 December 2018
in an optimal way, it is essential to know how much of the DNA in the sample remains in place and how much
Available online 23 December 2018
is transferred to the lifter. In the present study we addressed this question analyzing 16 pairs of thumb prints
and revealed that more than 80% of the DNA from a fingerprint is transferred to the gelatin lifter. Therefore,
Keywords:
subsequent DNA sampling of the stored gelatin lifters appears more promising than recovery of the residual
Forensic science
Fingerprints
DNA from the original fingerprint. Furthermore, as a proof of principle, we developed a protocol for the direct
DNA extraction of DNA from gelatin fingerprint lifters by proteolytic digestion of the gelatin matrix followed by
Gelatin lifters organic extraction. We show that DNA recovery from gelatin lifters by this direct extraction protocol is more
Extraction efficient compared to swabbing the lifter followed by standard magnetic bead extraction of swabs. However,
given the more elaborate protocol for direct extraction, we would still recommend the swab technique as the
method of choice for forensic routine work.
© 2018 Elsevier B.V. All rights reserved.

1. Introduction fingerprints, mainly focusing on the effect of fingerprint enhance-

Fingerprints and DNA traces are of eminent importance for
forensic investigations due to their potential for identification. It has
been demonstrated more than 20 years ago, that both types of traces
can be recovered from the same contact site [1]. In case work, latent
fingerprints are often visualized in situ by powder dusting. If the
mark is considered interpretable, it might either be photographed or
transferred to tapes, silicone or gelatin lifters for later examination
[2]. In most situations, examiners will not sample the same contact
spot for DNA if they get a good fingerprint. However, simultaneous
DNA sampling can be beneficial: if the person who left the mark has
not yet been registered, a DNA profile from the fingerprint might
permit to link DNA traces from different crime sites via trace-trace
database searching. The crime scene investigator should thereby
keep in mind the risk of cross contamination, when sampling at
different sites with the same material [3,4]. Since it has been shown
that sampling for DNA reduces the quality of latent fingerprints [5],
DNA is usually sampled after fingerprint development. Many studies
have been conducted on the simultaneous recovery of DNA and
ment techniques on the efficiency of DNA profiling, sampling the
enhanced fingerprint in situ [6–13]. There has also been some work
done on the recovery of DNA from lifted fingerprints, mostly
transferred to tapes [14–17], demonstrating their suitability for DNA
profiling. We could find two papers, involving widely used gelatin
lifters, demonstrating their suitability for DNA recovery and
subsequent profiling [18,19]. So far, no protocol has been published
for a direct extraction of DNA from gelatin lifters and the lifters are
usually swabbed for DNA. A direct extraction protocol has the
advantage that the whole DNA present on the lifter ends up in the
reaction tube, whereas with swabbing, DNA recovery from the lifter
will never be 100%. The present study serves as a proof of principle.
We developed a protocol for direct extraction of DNA from gelatin
lifters by proteolytic digestion and checked for DNA transfer rates
from fingerprints to gelatin lifters to get a better idea of what to
sample for DNA profiling: the stored fingerprint transferred to the
gelatin lifter, or the site where it has been lifted from?
2. Materials and methods

2.1. Sampling
* Corresponding author.
E-mail addresses: Martin.Zieger@irm.unibe.ch (M. Zieger),
Christophpaul.schneider@gmail.com (C. Schneider), Silvia.Utz@irm.unibe.ch Fingerprints for subsequent DNA sampling were taken from a
(S. Utz). male collaborator, not involved in the processing of the samples,

https://doi.org/10.1016/j.forsciint.2018.12.006
0379-0738/© 2018 Elsevier B.V. All rights reserved.
146 M. Zieger et al. / Forensic Science International 295 (2019) 145–149
without hand washing prior to deposition. The person mainly
carried out office work throughout a normal workday without any
further constraints. After rubbing his fingers against his neck or
forehead, the fingerprint donor pressed his thumbs onto a
previously cleaned (70% ethanol) glass plate. At least 30 min of
additional office work were carried out before the next sampling of
both thumbs. Since it has been postulated previously that black
powder might be more suitable for subsequent DNA processing
than metallic powders [10,13], fingerprints were visualized by
black powder (Sirchie1 Silk Black), applied by a fiberglass brush.
Fingerprints were covered by transparent BVDA1 gelatin lifters
(5  5 cm). Possible air bubbles were removed by sweeping a finger
over the backside of the lifter. The lifters were removed and the
remaining marks on the glass plate were taken up by a PrionicsTM
cotton swab (cardboard evidence collection kit; Thermo Fisher, US)
moistened with a drop of water from the vial included in the
collection kit. A total number of 16 thumb pairs and 8 untreated
controls were sampled. Samples were taken over four days,
resulting in 4 thumb pairs and two negative controls per day. The
sampling scheme is depicted in Fig. 1. Processing of left and right
thumbs with either method (direct extraction or swabbing) was
alternated for every pair of thumbs, to reduce a possible effect of
unequal DNA deposition between the dominant and non-domi-
nant hand.
2.2. Direct extraction
To reduce the amount of gelatin for digestion, the 16 powder
stained fingerprints (8 from left and right thumb each) were cut
out around the outlines of the mark from the 5  5 cm gelatin lifters
with a sterile disposable scalpel. From the 4 untreated control
lifters a 6 cm2 area was cut out, corresponding approximately to
the size of one of the thumb prints. The same scalpel was used to
remove the gelatin layer from the plastic support and transfer it
into a 2 ml tube. 500 ml of TNE (10 mM Tris–HCl, 100 mM NaCl,
10 mM EDTA, pH 8.0), 50 ml 1 M DTT, 50 ml 10% SDS and 50 ml
Proteinase K (20 mg/ml) (all compounds from Merck, Germany)
were added to the gelatin. The samples were shook on a
Precellys124 homogenizer (Bertin instruments, France) 2  30 s
at 5900 rpm followed by incubation over night at 56  C and
Fig. 1. Experiment setup: Both thumbs are pressed on a glass plate, visualized by black powder and lifted by gelatin lifters. One of the lifters is extracted directly and the other
one is swabbed. Residual DNA from both fingermarks is swabbed subsequently.
400 rpm on a thermoshaker. The next day additional 40 ml
Proteinase K (20 mg/ml) were added and the samples were
incubated another three hours at 56  C and 400 rpm. The reaction
volume was split into two equal volumes in two 2 ml tubes. 800 ml
of Phenol:Chloroform:Isoamyl Alcohol 25:24:1 (Sigma-Aldrich,
US) were added to each of the two tubes for extraction. The
aqueous phases were pooled in a Vivacon1 2 ETO column
(Vivaproducts, Inc., US) and cleaned by centrifugation at 2000 g,
successively adding 1 1 ml and 2  2 ml of distilled water to the
sample. The final elution volume was between 50 and 120 ml.
2.3. Swabs
The other 16 powder stained fingerprints on gelatin were
swabbed for DNA recovery by a Prionics cotton swab moistened as
described above. 4 swabs were taken from untreated gelatin lifters
as controls. All Swabs (from lifters and residual fingerprints) were
extracted with the AutoMateExpressTM device and the PrepFiler
ExpressTM Kit (both Thermo Fisher, US), as described in [20],
representing our standard lab procedure for swabs from touched
surfaces. Elution volume is 50 ml. Through an internal validation
study, we found that extraction of swabs by PrepFiler ExpressTM Kit
yields DNA amounts comparable to the ones obtained by organic
extraction. Even though this would have been suggested by the
organic extraction protocol for the lifters, we did intentionally not
choose an organic extraction protocol for swabs, because we aimed
to compare the efficiency of the whole workflow for gelatin lifters
with a realistic alternative workflow. Given the usually high
number of samples, organic extraction of standard swabs is not a
common procedure in most forensic labs.
2.4. DNA quantification and typing
DNA quantification was done by Real-Time-PCR (qPCR) using
the Quantifiler1 HP Kit from Life Technologies on a 7500 RT PCR
System (Thermo Fisher, US). DNA profiles from the collected
fingerprints and from the donor reference sample (buccal swab)
were established by multiplex-PCR using the AmpFlSTR1 NGM
SelectTM Kit (Thermo Fisher, US) in a total reaction volume of 25 ml
(single analysis). For amplification of samples with a DNA
 M. Zieger et al. / Forensic Science International 295 (2019) 145–149 147

concentration lower than 50 pg/ml, we used the maximum sample

volume of 10 ml, and for higher concentrated samples, we used
0.5 ng of DNA per reaction. In line with our standard operating
procedures for routine case work, all samples with a DNA
concentration below 20 pg/ml were amplified with 32 instead of
30 cycles. Capillary electrophoresis was run on a 3130xl genetic
analyzer (Thermo Fisher, US) and signal interpretation done with
Genemapper ID-X, v1.4 (Thermo Fisher, US). All peaks above 50rfu
were considered as true alleles.

3. Results

3.1. Direct DNA extraction from gelatin lifters

Detailed DNAyields from the gelatin lifters are shown in Fig. 2. The
numbers can also be found in Supplementary Table 1. In 13 out of 16
thumb pairs, more DNA was recovered by direct extraction than by
swabbing the lifter. We see that the DNA amount deposited varies
greatly from one sample to another, what could also explain the three
experiments for which the swabbing resulted in a better DNA yield.
Hence, the mean values for the extracted DNA in ng are 8.7 ng for
direct extraction and 4.7 ng for swabbing, with large 95% confidence
intervals (CI) of 5.2 ng and 5.4 ng, respectively. For the four
negative controls that have been swabbed, no DNA was detected by
qPCR. For the direct extraction, two out of four negative controls gave
a signal by qPCR, although indicating only a very low DNA
concentration (0.2 pg/ml and 0.4 pg/ml). With 10 ml input volume
and 32 PCR cycles, no STR-profiles could be established from those
two control samples. Interestingly and consistent with what has been
reported previously [21], we obtained in average more DNA from
thumbs of the non-dominant hand, even though this observation
proved not to be statistically significant (data not shown). In this
regard, it is noteworthy that for all three experiments where the
swabbing yielded more DNA than the direct extraction protocol, the
non-dominant thumb print has been analyzed by the swab protocol.
3.2. Comparison of direct extraction vs swab and transfer efficiency
Due to the large differences in DNA amounts deposited, the
average amounts of recovered DNA are not very meaningful.
Therefore, we normalized the values by using the fraction of DNA
extracted from the lifter, compared to the total DNA amount
Fig. 3. Average ratios of DNA detected on gel lifters over the total DNA amount (DNA
on lifters, as seen in Fig. 2 + residual DNA after lifting). n = 16 samples were analyzed
per method. Direct digestion is significantly more efficient with *p = 0.04 (two-sided
Students t-test). see Supplementary Table 1 for detailed DNA amounts.
deposited with every thumbprint (Fig. 3). In average we could
detect 85% of the total DNA on the lifter for direct extraction,
compared to a recovery of 67% for the swabbed lifters. Since the
sampling is the same for all fingerprints, this difference must be
due to the different extraction protocols. Despite the small number
of samples used for this study, the difference is statistically
significant with p = 0.04 (two-sided paired t-test). All DNA amounts
are given in Supplementary Table 1.
3.3. Quality of profiles
Autosomal 16-STR profiles (AmpFlSTR1 NGM SelectTM) were
established from every sample. We counted the number of loci for
which we obtained the allele values of our donor profile.
Heterozygote loci passed if the sister peak height was at least 60%.
The results can be seen in Fig. 4. We did not take special precautions
to prevent contamination, since this was not the relevant issue
within the scope of this study. However, interpretable DNA profiles
originated exclusively from the donor. One of the directly extracted

Fig. 2. DNA amounts sampled from gelatin lifters by either swabbing or direct extraction. One experiment (Exp) stands for one pair of thumbs. Four experiments (01–04, 05–
08, 09–12 and 13–16) were conducted on four different sampling days consecutively. For odd-numbered experiments, the thumb prints from the non-dominant hand were
extracted directly, for even-numbered experiments, the thumb prints from the dominant hand were extracted directly. see Supplementary Table 1 for detailed DNA amounts.
148 M. Zieger et al. / Forensic Science International 295 (2019) 145–149
Fig. 4. Average number of typed loci, suitable for database submission. The residual
DNA amounts from fingerprints lifted and directly extracted by digestion (“Residual
direct”) and from fingerprints lifted and swabbed from the lifters (“Residual swab”)
are shown separately. Error bars indicate standard deviations.
lifter samples apparently contained PCR inhibitors. Despite a high
DNA concentration of 0.14 ng/ml indicated by qPCR, no interpretable
STR profile could be established. After an additional purification step
of this sample with PrepFiler ExpressTM, a complete profile of good
quality was obtained.
4. Discussion
We established a simple protocol to directly extract DNA from
gelatin fingerprint lifters. Our results show that we detect a higher
percentage of the total DNA from a fingerprint on the gelatin lifters
if we use direct extraction followed by organic extraction,
compared to swabbing the lifters followed by magnetic bead
extraction of the swabs. Since both fingerprints are always lifted
the same way, this difference must be due to the different
subsequent processing of the lifters. Thus we conclude that DNA
recovery by the direct proteolytic digestion protocol is more
efficient. The approximately 25% increase (Fig. 3) in DNA obtained
by the direct extraction protocol could be decisive if the DNA
content of a trace is close to the detection threshold. This might be
particularly interesting for high-profile cases.
However, for the fresh and DNA-rich fingerprints in this study,
the difference is negligible since both methods led to a comparable
number of typed loci. The small positive effect together with the
much more time consuming protocol for direct extraction, leads us
to the conclusion that swabbing the lifter is still the method of
choice for samples from volume crimes.
If one has to choose, we recommend analyzing the DNA on the
lifter and not the residual DNA. Our results clearly demonstrate
that most of the DNA material (more than 80%) from the
fingerprint is transferred to the gelatin lifter. For the setup we
chose here, the choice of the sampling site (lifter or residual) did
not make a significant difference in terms of interpretable DNA
profiles, since it was also possible to obtain good profiles from the
residual DNA. But again, it might become crucial for traces
containing small amounts of DNA.
We did not check the uptake of possible PCR inhibitors by the
gelatin lifters from the sampling site. All prints were taken from
pre-cleaned surfaces. However, we would not expect enrichment
of inhibitors on the lifters, compared to DNA sampling by swab. The
study focused on fingerprints on glass surfaces. We would expect
similar results for other plane and smooth surface, but transfer of
DNA to the fingerprint lifter might be different for rough surfaces.
For labs interested in establishing the present protocol, in-house
validation of other combinations of powder, gelatin and swab, as
well as comparison to established extraction protocols is
recommended.
Without further validation, the direct extraction protocol
should be used exclusively for transparent gel lifters from BVDA1,
since we noticed that the composition of the lifter can have an
impact on the quality of the DNA extraction. In preliminary
experiments, we were not able to extract high quality DNA by
direct digestion of BVDA’s black and white gelatin lifters (data not
shown). So we assume that those lifter types might contain
inhibitors such as metal compounds, acting as coloring substances.
Those inhibitors could probably be removed by an additional
purification step. Yet this would lower the DNA yield and therefore
probably render the direct extraction protocol useless. Herewith
we would also like to encourage the manufacturers of gelatin lifters
to consider the production of an inhibitor-free and DNA-free
gelatin lifter, optimized for direct extraction. We were surprised
however, that almost no contaminating DNA was detected on the
lifters even though they are not explicitly manufactured to be DNA-
free. Only some minor alleles were detected in a couple of samples,
originating from unknown contributors. Thus, a contribution of
extraneous DNA is also unlikely to have an impact on the outcome
of our study in terms of DNA quantity. Nevertheless, for real case
work samples – also given the potential risk of cross contamination
by brushes used for powdering – we strongly recommend to either
use DNA profiles from fingerprints only as intelligence tool, with an
awareness of possible contamination [6] or to use disposable
brushes, respectively clean the brush, as has been described
recently [4].
5. Conclusion
We could demonstrate that upon fingerprint sampling, most of
the DNA from the fingerprint is transferred to the gelatin lifter. As a
proof of principle, a method for direct extraction of DNA from
transparent gelatin lifters has been presented.
CRediT authorship contribution statement
Martin Zieger: Conceptualization, Methodology, Formal anal-
ysis, Writing - original draft, Writing - review & editing. Christoph
Schneider: Investigation. Silvia Utz: Resources.
Acknowledgements
We thank Britta Stoop and Colin Tièche for critical reading,
Sonja Burri for technical assistance, David Comment from the
Bern Cantonal Police for practical advice on fingerprint sampling
and BVDA1 for providing us with free samples of gelatin lifters.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at https://doi.org/10.1016/j.forsciint.2018.12.006.
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