Forensic Science International: Genetics Supplement Series xxx (xxxx) xxx–xxx

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Forensic Science International: Genetics Supplement Series

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

Materials and methods that allow fingerprint analysis and DNA profiling
from the same latent evidence
⁎
Alexander Sinelnikov , Karl Reich
Independent Forensics, Lombard, IL, USA

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

Keywords: The analysis of forensic evidence for both latent ridge impressions (fingerprints) and DNA profiling has long
Latent fingerprints been considered mutually exclusive; processing and sensitivity issues precluded this dual analysis.
Touch DNA By systematically reexamining the techniques and reagents used by these forensic identification methods, we
Hinge cards have developed simple, practical solutions that provide latent examiners with the high-resolution image they
Subtractive spin-column DNA purification
require and facilitate the recovery of the minute amount DNA in a fingerprint for successful DNA profiling by the
Post-PCR purification/concentration
DNA STR profile
forensic laboratory.

1. Introduction
Previous research in developing DNA STR profiles from individual
fingerprints noted the poor recovery of DNA from this type of evidence
[1]. This is primarily due to the limiting amount of DNA in an in-
dividual fingerprint and the losses incurred during DNA processing and
purification steps. Some researchers have focused on using cell-free
DNA and direct amplification which avoids the losses during extraction
and purification [2]; however this approach limits the types of evidence
that can be processed. Other researchers have used blood or saliva
produced fingerprints where of course the amount of DNA is not lim-
iting [3]. Here we use “skin and oil” fingerprints identified with pow-
ders and ‘lifted’ onto sticky tape or hinged cards which actually mimic
real world evidence and currently used techniques.
2. Materials and methods
2.1. Sample collection
Individual skin and oil fingerprints identified with DNA-free black
powder (Sirchie, Youngsville, NC) were lifted onto adhesive surfaces of
hinge cards and closed with a transparent liner (IFI, Lombard, IL;
Sirchie, Youngsville, NC) (Fig. 1A, B, D). The design of this lift card
permits easy access to the adhesive-lifted fingerprint after it has been
imaged by the latent examiner. Biological material left on the evidence
surface after lifting (Fig. 1C) was collected using a sterile cotton swab
(Puritan, Guilford, ME) moistened with 10 μL collection buffer (10 mM
Tris–HCl, 0.1 mM EDTA, 0.25% SDS) (Fig. 1E). This swab was reserved
and subsequently used for collecting the liquid extract derived from the
enzymatic digestion of adhesive layer of the lift card within the lysis
chamber (Fig. 1G).
2.2. DNA extraction
DNA extraction (proteolytic digestion in lysis buffer: 10 mM
Tris–HCl, 0.1 mM EDTA, 0.25% SDS, 100 μg/mL Proteinase K) was
performed for 1 h at 56 °C directly on the adhesive layer of lift cards
immobilized in the lysis chamber (Fig. 1G) and on the spin-basket
collected material derived from the reserved swab (i.e., recovered evi-
dence not lifted by the adhesive card) (Fig. 1E and I). Lysis chamber
assembly consisted of retaining the hinge card, adhesive layer upper-
most, on the designated center of the chamber, covering the exposed
adhesive with derivatized glass fiber (to convert the hydrophobic ad-
hesive to a hydrophilic surface), placing the provided O-ring on the
bottom of the chamber, adding the lysis/digestion buffer to the lifted
fingerprint and then adding the second glass slide, producing an air and
moisture-tight chamber that is compressed and sealed with two binder
clips (Fig. 1G).
2.3. DNA purification
The Xs DNA purification spin column (IFI, Lombard, IL) uses a resin
that retains proteins, SDS, DTT, ninhydrin, and fingerprint powders, but
does not bind DNA (Fig. 1J). Thus, DNA passes through the resin while
a range of PCR inhibitors are retained on the column. The column is
prepared by an initial centrifugation, 5 min, 14,000 RCF, digestion

⁎
Corresponding author at: Independent Forensics, 500 Waters Edge, Suite 210, Lombard, IL 60148, USA.
E-mail address: alex@ifi-test.com (A. Sinelnikov).

http://dx.doi.org/10.1016/j.fsigss.2017.09.010
Received 12 August 2017; Accepted 10 September 2017
1875-1768/ © 2017 Elsevier B.V. All rights reserved.

Please cite this article as: Sinelnikov, A., Forensic Science International: Genetics Supplement Series (2017),
http://dx.doi.org/10.1016/j.fsigss.2017.09.010
A. Sinelnikov, K. Reich Forensic Science International: Genetics Supplement Series xxx (xxxx) xxx–xxx

Fig. 1. Work flow schematic: DNA profiles from latent ridge impressions.
A) Starting material: latent ridge impression enhanced with powder (fingerprint). B) Lifted fingerprint on hinge card (suitable for imaging). C) ‘Ghost’ fingerprint left on evidence post-
lifting. D) Exposed fingerprint on lift card, note transparent liner being held by gloved hand. E) Collection of ‘left-behind’ material using swab and collection buffer. F) Represents total of
fingerprint evidence = lift card + recovered left-behind material. G) Immobilized lift card in lysis chamber, note O-ring, binder clips. H) Swab used to recover left-behind material is used
to collect extract from incubated lysis chamber. I) total lysate recovered after spin basket; note that a second proteolytic digestion is performed on this material. J) Combined lysa-
te = lysate from lift card + lysate from reserved swab purified on Xs column; note retained powder in column (left) and clear purified lysate (right). K) GeneScan electropherogram of CE
analyzed PowerPlex 16 PCR reaction.

extract is added to the top of the spin column and purified DNA is re-
covered after a second centrifugation, 5 min, 8000 RCF.
2.4. PCR amplification
Identifiler (ABI, Waltham, MA) and PowerPlex 16 (Promega,
Madison, WI) PCR reactions were prepared as per manufacturer’s in-
structions and amplified for 29 cycles (Identifiler) and 32 cycles
(PowerPlex 16).
2.5. Post-PCR purification and capillary electrophoresis
Samples that generated partial profiles were subjected to post-PCR
purification/concentration using AmpliconRx™ (IFI, Lombard, IL) [4].
Briefly, PCR reactions were prepared by the addition of binding buffer,
binding the amplicons to the spin column with purification facilitated
by discarding the column run-through (i.e., nucleotides, unincorporated
primers, salts, enzymes and buffer) and eluting amplicons with for-
mamide/size standards mixture [4]. The full volume (∼15 μL) of the
eluate was used for capillary electrophoresis on a ABI3100 instrument
(ABI, Waltham, MA).
3. Results and discussion
The overall procedure we have developed is typical for a forensic
protocol: evidence collection on swabs, proteolytic digestion, DNA
purification, amplification and analysis. The changes to the current
procedures increase efficiency of sample collection, DNA purification
and post-PCR steps as described below.
(1) By using detergent-based collection buffer, as opposed to water,
there is a substantial improvement in collection efficiency, in re-
ducing the volume of liquid needed per surface area and better
recovery of DNA from the swab during spin-basket centrifugation
[5].
(2) By collecting the enhanced ridge detail that is not lifted by the
adhesive hinge card, we can recover up to 50% more biological
material than from the hinge card alone. The latent examiner may
have his/her pattern preserved on the hinge card, but the DNA
analyst needs all of the biological material = hinge card + the
material left behind.
(3) By using the ‘subtractive’ purification approach (i.e., a column that
binds PCR inhibitors including detergent, proteins and DTT, but

2
A. Sinelnikov, K. Reich
allows DNA to pass through the resin), we have improved the re-
covery of DNA from fingerprint evidence. Our estimate of DNA
recovery efficiency is based on DNA-STR profile quality using
known amounts of purified DNA on Xs columns and then scoring
the resulting electropherograms. Average DNA recovery from Xs
column purification, 16 replicates, was 93%.
(4) By developing a post-PCR purification/concentration kit, we have
increased capillary electrophoresis signal 5–10-fold for 12.5 μL PCR
reaction and 10–20-fold for 25 μL PCR reaction [4]. Current post-
PCR procedures only analyze 4–8% of the PCR reaction (1 μL from
either a 12.5 or a 25 μL PCR reaction). By purifying the amplicons
away from electrokinetic injection inhibitors (salts, unincorporated
PCR primers and nucleotides) far more amplified fragments enter
the capillary with a concomitant increase in CE signal.
The complete procedure was tested on twelve individual finger-
prints which were deposited on non-porous surfaces, identified with
black fingerprint powder, lifted onto our hinge card (and the left-be-
hind material collected as well) and extracted/purified as described
(see Section Materials and methods). The DNA profiles from these
samples demonstrated recovery, on average, 80% of expected alleles
(Identifiler and PowerPlex 16 PCR amplification kits).
This procedure was also tested on latent fingerprints identified with
powders but not lifted onto adhesive cards mimicking fingerprint evi-
dence deemed not suitable for comparison; ten examples: five finger-
prints identified with black powder and five fingerprints identified by
fuming (vaporized cyanoacrylate) and Rhodamine 6G staining. These
samples were individually collected and processed (see Section
Materials and methods), but without using the lysis chamber. The DNA
profiles from these samples demonstrated recovery, on average, 92% of
expected alleles (Identifiler PCR amplification kit).
In a companion study the ‘subtractive’ DNA purification and post-
PCR processing approach were used on laser captured sperm cells; here
processing 25 counted sperm cells (i.e., 12.5 diploid cells) demonstrated
an average allele recovery of 87% with a 24-locus multiplex (PowerPlex
Fusion), thus illustrating the sensitivity of this approach and its
Forensic Science International: Genetics Supplement Series xxx (xxxx) xxx–xxx
flexibility (i.e. applicability to other types of low template amount
samples).
4. Conclusion
By developing methods and reagents specifically for evidence with
limiting and minimal amounts of DNA, we have been able to increase
the sensitivity of DNA profiling such that the smallest individualized
item of evidence, a single fingerprint, can now be processed and ana-
lyzed by forensic DNA laboratory. The method can also be applied to
other evidence with limiting amounts of DNA.
Conflict of interest
Other than being employees of Independent Forensics, the authors
have no conflict of interest.
Acknowledgements
The authors would like to thank Raymond J. Miller, Jr., CLPE (US
Custom and Border Protection Southwest Regional Science Center,
Houston, TX) for samples of fingerprints identified by vacuum fuming
with cyanoacrylate and stained with Rhodamine 6G dye.
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