42(3):225-228,2001

FORUM

Preservation and Collection of Biological Evidence

Henry C. Lee, Carll Ladd

Forensic Science Laboratory, Division of Scientific Services, Connecticut Department of Public Safety, Meriden,
Conn, USA

As the courts have placed greater emphasis on physical evidence during the past few decades, the initial stages of evi-
dence examination have become increasingly important to the successful resolution of many criminal investigations.
This emphasis on evidence collection and preservation is often manifested by many rigorous court challenges. This ar-
ticle reviews how the ability to introduce DNA test results in court is affected by methods used to recognize, document,
collect, and preserve biological evidence.
Key words: base sequence; DNA; expert testimony; forensic medicine; genes; preservation, biological

During the past few decades, physical evidence The quantity of DNA that can be extracted from
has become increasingly important in criminal inves- these common biological sources will vary (Table 1).
tigations. Courts often view eyewitness accounts as Note that, in practice, crime scenes samples may con-
unreliable or biased. Physical evidence, such as tain considerably less usable DNA depending on envi-
DNA, fingerprints, and trace evidence may independ- ronmental conditions. DNA has been isolated from
ently and objectively link a suspect/victim to a crime, other sources, such as gastric fluids and fecal stains.
disprove an alibi, or develop important investigative However, it can be difficult to generate a DNA profile
leads. Physical evidence may, also, prove invaluable from these sources in case samples due to significant
for exonerating the innocent. degradation.
The initial stages of physical evidence examina- Several factors affect the ability to obtain a DNA
tion can be pivotal to the successful resolution of profile. The first issue is sample quantity. The sensitiv-
criminal investigations. The methods employed in the ity of polymerase chain reaction- based (PCR) DNA
recognition, collection, and preservation of physical typing methods is noteworthy, but still limited. The
evidence, such as DNA, have been rigorously scruti- second concern is sample degradation. Prolonged ex-
nized and challenged in court. posure of even a large blood stain to the environment
or to bacterial contamination can degrade the DNA
Sources of DNA and render it unsuitable for further analysis. The third
consideration is sample purity. Most DNA typing
The forensic application of DNA typing methods methods are robust, and dirt, grease, some dyes in
over the past fifteen years constitutes a major ad- fabrics, and other substances can seriously compro-
vancement in the examination of biological evi-
dence. With its remarkable sensitivity and power of Table 1. DNA content of biological samplesa
discrimination, DNA analysis has become a key fig- Type of sample Amount of DNA
ure in the fields of forensic science, forensic medicine Liquid blood 20,000-40,000 ng/mL
and anthropology, and paternity testing (1). stain 250-500 ng/cm2
Many different types of physical evidence are Liquid semen 150,000-300,000 ng/mL
commonly submitted to forensic science laboratories Postcoital vaginal swab 10-3,000 ng/swab
for examination. Initially, evidence that was suitable Hair (with root)
for DNA analysis was limited to biological substances Plucked 1-750 ng/root
that contain nucleated cells. This limitation has been Shed 1-10 ng/root
overcome in the last 5 years with the implementation Liquid saliva 1,000-10,000 ng/mL
of mitochondrial DNA sequencing in the forensic Oral swab 100-1500 ng/swab
arena. Common biological specimens from which Urine 1-20 ng/mL
DNA has been successfully isolated and typed are as Bone 3-10 ng/mg
follows: bones, blood and bloodstains, semen and Tissue 50-500 ng/mg
seminal stains, tissues, organs, teeth, hairs, finger- a
Quantity of DNA recovered from evidentiary samples is significantly
nails, saliva, urine, and other biological fluids. affected by environmental factors.

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mise the DNA typing process. Environmental insults be processed or moved until its original condition
will not change DNA allele “A” into allele “B”, but and other relevant information have been recorded.
they can adversely affect the ability of the scientist to Several different means of documentation are avail-
obtain a complete DNA profile from the sample (2-4). able. Generally, the use of more than one method is
advised. The basic approach of evidence documenta-
Evidence Transfer tion and handling is outlined in Tables 2 and 3.
DNA evidence can be used to make linkages or
associations (e.g. person-person, person-other physical Collection and Preservation of Biological
evidence, or person-crime scene). In general, biologi- Evidence
cal evidence can be transferred by direct deposit or The ability to introduce DNA findings in court is
by secondary transfer. also greatly impacted by evidence collection and
preservation methods. Evidence integrity, both scien-
Direct Deposit tific and legal, begins with the first investigator at the
Any biological evidence (blood, semen, body tis- crime scene. Detailed evidence collection protocols
sue, bone, hair, urine, and saliva) can be transferred have been previously described (7-13). The specific
to an individual’s body/clothing, object, or crime collection method employed will depend on the state
scene by direct deposit. Once biological fluids are de- and condition of the biological evidence. In general,
posited, they adhere to the surface and become a significant quantity of material should be collected
stains. Non-fluid biological evidence, such as tissue to ensure the recovery of sufficient DNA for testing
or hair, can also be transferred by direct contact. purposes. However, it is important to limit collecting
Secondary Transfer additional dirt, grease, fluids, and other material from
the surrounding area, since many substances are
Blood, semen, tissue, hair, saliva, or urine can be known to adversely affect the DNA typing process.
transferred to a person, object, or location through an Each biological specimen should be packaged ac-
intermediary (person or an object). With secondary cording to established forensic practices. Once the
transfer, there is no direct contact between the origi- samples have been collected, they should be
nal source (donor of the biological evidence) and the promptly delivered to the forensic laboratory. To min-
target surface. Secondary transfer may, but does not imize specimen deterioration, items should be stored
necessarily, establish a direct link between an indi- in a cool, dry environment until they are submitted
vidual and a crime. The impact of secondary transfer for testing.
on the interpretation of DNA results has been debated
(5,6). However, secondary transfer is clearly a more Many famous investigations, such as O.J.
significant concern with the more sensitive DNA typ- Simpson and J.B. Ramsey, highlight the importance of
ing methods, such as mitochondrial DNA sequencing effective crime scene processing (13,14). In the legal
and low copy number PCR. arena, unless the evidence is properly documented,
collected, packaged, and preserved, it may not meet
the legal and scientific requirements for admissibility
Evidence Recognition into a court of law. If the DNA evidence is not prop-
The first step in a criminal investigation is deter- erly documented before the collection, its origin can
mining which samples warrant further testing. This be questioned. If it is improperly collected or pack-
phase is crucial to the outcome of the investigation aged, the possibility of contamination will be raised
and very challenging, as crime scenes can be both to discredit the DNA results. Given the prospect of le-
complex and chaotic. Hence, an experienced investi- gal challenges and the sensitivity of PCR methods, it
gator who systematically evaluates the scene is an in- is essential that strict contamination prevention mea-
valuable resource. Recognition is the ability to iden- sures be followed.
tify probative evidence (at the scene or in the labora- Legal concerns often diverge from empirical
tory) scattered among potentially vast quantities of re- data. Even though PCR-based typing methods are sen-
dundant, irrelevant, or unrelated items. For instance, sitive, the contamination argument has been exagger-
collecting 20 bloodstains from the vicinity of a stab- ated in some cases (5,15). Moreover, it is important to
bing victim may not point to the perpetrator. The rec- note that, since all multi-locus DNA profiles (ie, >6
ognition process involves basic forensic principles, STR loci typed) are rare, contamination will predomi-
such as pattern recognition and analysis and physical nantly lead to false exclusions or artificial mixtures
properties observation. Naturally, if crucial evidence rather than false inclusions. Consequently, albeit con-
is not recognized, collected, and preserved, its value tamination could complicate result interpretation, it
to the trier of fact will be lost. would typically not include the defendant.

Documentation of DNA Evidence Challenges to DNA Admissibility

The location and condition of any biological evi- Since their introduction into forensic science,
dence must be thoroughly documented before its col- DNA typing methods have been strenuously attacked
lection. Careful evidence documentation at the crime in court (Table 4). Initially, the reliability of DNA typ-
scene, autopsy room, and forensic laboratory is es- ing procedures was questioned along with the statisti-
sential. In any criminal or civil investigation, docu- cal methods used to calculate DNA profile frequen-
mentation has great bearing on whether the evidence cies. In the last few years, legal challenges regarding
can later be introduced in court. Evidence should not the admissibility of DNA have shifted their focus

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away from the general reliability of the methods. Al- quent handling of the biological evidence. The spec-
though most courts are comfortable with DNA testing ter of evidence tampering may also be raised. An-
in principle, some defense objections regarding DNA other common case-specific challenge concedes that
evidence continue to be effective. Successful chal- DNA typing methods are reliable in theory. Here, the
lenges to the admissibility of DNA testing often ad- defense may suggest that critical mistakes were made
dress the initial collection, preservation, and subse- in testing (sample switches, contamination, devia-

Table 2. Evidence documentation and collection

A. Evidence at a crime scene
Photograph the evidence before it is touched, moved, or collected.
Videotape the evidence and its relative position at the crime scene.
Document the location and condition of the evidence.
Note and sketch the spatial relationship of the evidence relative to other objects at the scene.
Label, initial, and seal the evidence package.
B. Evidence at the forensic laboratory
Note the package, label, and seal condition of the item.
Label the package with initials, unique case identifier, and date.
Check the item number and compare it to the submission form to ensure that the correct item has been received. Also verify that
the description of the item is accurate.
Note, sketch, and/or photograph the contents of the package.
Document the location and condition of biological evidence on the item prior to any sampling. Note when secondary cuttings of
the evidence are taken; include the area where the cutting was collected. Package any sub-items separately.
When testing is conducted, record quantity of sample consumed, the test performed, and the results obtained. When handling the
evidence, always wear clean, disposable gloves to minimize contamination.
C. Evidence at the autopsy room
Photograph the body and any additional evidence before cleaning the body.
Note and sketch the evidence.
Systematically collect each piece of evidence with clean tools.
Separately package each item in a proper container.
Label the container and note the quantity of sample collected. Do not add preservatives, such as formaldehyde to the specimen.
Store the item appropriately.
Carefully collect the clothing to avoid losing trace evidence and to avoid contamination with other biological samples.
Release the evidence according to proper procedures.

Table 3. Laboratory processing of DNA evidence

A. Laboratory receipt of evidence
Physical evidence should be submitted to the laboratory with a transmittal letter, inventory sheet, and notation of the type of
examination requested for each item according to standard laboratory protocols.
All identifying information on the physical evidence should be checked against the submission forms. Any discrepancies should be
noted and corrected.
Each package should be properly packaged, sealed, and labeled. Any sign of improper packaging, sealing, or labeling should be
noted.
Note any sign of sample leakage or contamination.
Any special requests/instructions regarding the DNA testing should be recorded on the submission form.
A receipt for evidence showing the date, time, submitting agency, submitter’s name, case number, item numbers, and the
receiver’s name should be issued.
Physical evidence submitted for DNA analysis should be transmitted as soon as possible to the DNA unit and stored appropriately.
B. Laboratory initial processing procedures
An evidence examination form should be used to record the preliminary processing of each item. It should contain the following
data:
a) Package description and actual contents;
b) Label information, local case number;
c) Description/condition of evidence;
d) Laboratory case and item number;
e) Date and initials of examiner.
Document the size, location, pattern, and condition of the stained area.
Weigh biological evidence, such as bone, teeth, nail, and tissue as necessary. Note the quantity used for DNA analysis.
Record any preliminary test results and any trace evidence recovered.
Record information about each sample subjected to DNA analysis, as follows:
Case number, item number, and description; examiners’ initials;
Reagent lot number; protocol followed; quantity of sample consumed.
Testing results on each item should be entered on the appropriate worksheets.
Handle samples carefully to avoid mislabeling or cross contamination.
Whenever feasible, a portion of the sample should be preserved for possible future analysis. These specimens should be stored in a
freezer. However, in many instances the item cannot be divided due to insufficient quantity. In this event, the sample should be
processed according to standard forensic laboratory guidelines.
Any secondary cutting for DNA analysis should be placed in a separate container, package, or tube (labeled accordingly).
Unused DNA should be properly labeled and stored in a freezer.

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