Preface

What follows is intended to be a brief

review of the techniques employed by the
ILE to collect, analyze and interpret DNA
analysis results. We have not seen the
Kercher’s case actual lab reports nor were
we present when the samples were
collected and successively stored, but we
trust the judgment of several judges, and
the reports of professional and well-
reputed scientists working on the case
who have stated international standard
guidelines have always been followed in
the stages of sample collection, storage,
and DNA analysis.
 DNA collection, profiling
and interpretation: the ILE
at work
Content:

-Suitable biological samples

-Italian Forensic analysts: who are they, how do they

collect samples, what DNA analysis protocols do they use?

-Interpretation of Results
 Step 1: collecting
samples
What are the biological samples
more likely to be found on a crime
scene?
Human DNA can be extracted from any
fluid or bodily (saliva, blood, semen,
hair, nails etc) It can also be extracted
from any object that has been
contaminated by any bodily fluid such
as chewing gum, cigarette butts,
fabric, metals, paper tissues etc.
SAMPLES FROM WHICH DNA CAN BE EXTRACTED
• blood
• sperm
• organic tissues
• saliva
• sweat
• bones
• teeth
• urine
• feces
Can DNA samples be contaminated during collection
phase?

Yes, they can. DNA can be contaminated by external

sources, but since DNA DOES NOT FLY AND NEITHER
IT WALKS, it can only be transferred passively.

In order to understand the dynamics of

passive DNA transfer, let’s consider two
possible scenarios…
 …Scenario no. 1 or THE
INCOMPETENT COP
Let’s say my neighbor gets murdered (God
forbid!!). The cops knock at my door to ask me if I
heard something. I let them in, and shake hands.
The cats come by and one cat-lover cop stroke
them. They enter my neighbor apartment and
start collecting evidence wearing no gloves or no
shoe-covers….
…They find traces of my DNA and cats hair on
several pieces of evidence collected at the crime
scene, including my neighbor bra and other items I
had no business touching when paying friendly
neighbor visits in the past. Although this wouldn’t
rule me out as a suspect-unless of course I have a
sound alibi-it could be a case of DNA contamination.
Scenario no. 2 or THE COMPETENT
COP
Let’s say my neighbor gets murdered (God forbid!!). The
cops knock at my door to ask me if I heard something. I let
them in, and shake hands. The cats come by and one cat-
lover cop stroke them. They enter my neighbor
apartment…
…After they have carefully covered their shoes with
shoe covers and wearing gloves that are changed
every time they handle a new piece of evidence

Do they find traces of my DNA and cats hair on some

pieces of evidence collected at the crime scene?
Perhaps, after all I was a frequent visitor there-assuming
no bleaching has been performed-But do they find traces
of my DNA on my neighbor bra or any other item I had
no business to touch when paying friendly neighbor visits
in the past? Very unlikely.
Unless someone can show the opposite
occurred, we can say that correct sample
collection techniques have been used at
the crime scene in Via della Pergola. DNA
is not a virus or a bacterium which can be
transferred by breathing on objects.
Contamination of the DNA specimens
gathered at the murder scene was
therefore extremely unlikely in the
collection phase.
Who are the Scientific Police?
 The Polizia Scientifica as well as the RIS –
Reparto Investigazioni Scientifiche of the
Carabinieri- are cops holding University
and graduate degrees in biology,
chemistry and physics, among others.
 Many also hold a Master degree in
Forensic Sciences.
 The ILE operates according to the
recommendations of the DNA Commission
of the International Society for Forensic
Genetics.
 Storage of samples  

All samples (blood, saliva, tissues,

objects etc) can be stored at room
temperature for 24 hours. For longer
periods the samples must be stored at
– 20 C except for sample slides
preparations, FABRICS AND OBJECTS
that may continue to be stored at
room temperature.
So, was the bra clasp contaminated
or the DNA degraded in any way
while sitting in sealed crime scene
for a few weeks?

Very unlikely
Analysis results interpretation according to
the SIGU recommendations, following
COUNCIL OF EUROPE
Committee Recommendation No. R (92) 1
Genetic profiling results can lead to:
Exclusion: the DNA cannot be traced
back to the suspect
Compatibility: only a partial
analysis can be performed on
“difficult substrates; identity cannot
be ruled out but it is not attributable
with high probability
Identity: material present in
discrete/good quality/quantity.
Probability of having same identity
extremely high.
 PCR analysis
 Contamination during this stage may
always occur in any lab. However, several
protocols exist that help to minimize the
danger.
 The biologist in charge has assured that
maximum care was employed to avoid any
chance of contamination.
 However, test are always repeated just to
make sure that results are not false
positive (or even false negative)
Has the defendants’ DNA been found non-
compatible with the traces found on the
victim body/crime scene?
NO
Has the defendants’ DNA been found
compatible with the traces found on the
victim body/crime scene?
NO
Has the identity of the defendants’ been
established as per their DNA found on the
victim body/crime scene?

YES
 Theanalysis from 9 a 15 STR allows
to establish a probability of match
over 99,9999%, leading to a report of
identity as signed by the biologist in
charge of the analyses.
 STRs used in human DNA fingerprinting
ENFSI CODIS
(7 loci) (13 loci)
TH01 TH01
D21S11 D21S11
D18S51 D18S51
vWA vWA
FGA FGA
D8S1179 D8S1179
  TPOX
  CSF1PO
  D16S539
  D7S820
  D13S317
  D5S818
D3S1358 D3S1358

ENFSI (European Network of Forensic Science

Institutes)
CODIS (Combinated DNA Index System) della FBI. 
Individuals differ from one another in
terms of their STR profile.

Currently, fifteen markers are being

used in the determination of genetic
profiles
 Problems
Given that during collection, handling
and performing of the DNA analysis
proper procedures have been
followed as stated by several
authoritative sources, sometimes
interpretation problems may occur
such as…
Unclear peaks

 Unclear peaks due to

technical problems;
these peaks may be
mistakenly interpreted
as alleles
 Possible Problems when DNA is
scarce or degraded (Allele “dropout)
Reference sample

1500

Sample to be analyzed

150

?
Sometimes only one of the two alleles at a locus is amplified. In the
above figure, allele 14 at site D13S317 hasn’t amplified
 However, it is well known how
they are part of the most
common defense strategies,
such as :
• Contamination
• Statistical Weight of a Match
• Degradation/PCR Inhibition of “True”
Perp
• Artifacts (N+4 stutter, etc.)
• Thresholds Set Too High (missing
peaks)
• Examiner Bias
We know at times some of it may
happen…
…however the evidence
produced by prosecutors
should be argued on the base
of scientific expert knowledge
and not by reasoning that…
the crime took place in Italy
hence the results-curiously
limited to two of the
 References
 Hammond et al, 1994 Am J Hum Genet
55:175-189;
 Garofano et al, 1998 J Forensic Sci 43: 837-
840;
 Garofano et al., 1998 Forensic Science Int.
97: 53-60)
 www.bioforensic.com/conference07/index
 John M. Butler "Forensic DNA typing:
biology and technology behind STR
markers".1st ed ACADEMIC PRESS 2001
 N. Rudin and K. Inman,“An introduction to
Forensic DNA analysis”Second edition CRC
Press 2002