BIOLOGICAL EVIDENCE

COLLECTION
FOR

DNA EXAMINATION

DR.I.HAQUE
CFSL,KOLKATA
 FORENSIC APPLICATIONS OF
DNA FINGERPRINTING

  Murder/Attempt to murder
  Rape/Sodomy/other sexual offences
  Accidents/mass Disaster
Identification of mutilated bodies
Fake encounters/ Identification of
Skeletal Remains
  Parentage Disputes/ Baby Sweeping
Cases/ Missing Identity
Burglary/Threatening/Crime Scene
Investigation etc
Evidence Collection

 Collection of Evidence

Types of Unknown Samples:

 Blood, Semen, Stains, Saliva
 Hair, Tissue, Bones, Teeth

Types of Known Samples:

 Blood or buccal swabs from suspect
or victim or other known person
 GENERAL GUIDE LINES FOR INVESTIGATION OFFICER:

Collect and Package Physical Evidence – physical evidence must

be handled and packaged in a way that prevents any change from
occurring. The evidence must be properly packaged according to
type and properly labeled and sealed with appropriate initials to
maintain chain of custody. The integrity of evidence is best
maintained when it is kept in its original condition as found at the
crime scene. Trace evidence should not be removed from the
objects they’re found on unless it is impossible to transport the item.
1) Proper Packaging – The crime scene investigator
should bring several different types of packaging with them to the
scene. Small packages to contain hairs, fibers, bullets, glass or
other items should be available typical types of small packaging
include:
a) screw-cap or pressure-lid plastic bottles
b) coin envelopes/manila envelopes
c) small pill boxes
d) “druggist fold” paper bindles
GENERAL GUIDE LINES FOR INVESTIGATION OFFICER:

Collect and Package Physical Evidence

1) Proper Packaging (cont’d) – While these types of

sealable packages are good for powders and other
dry trace, bloodstains and bloodstained
evidence is different. Bloodstained evidence
should not be packaged in airtight containers due to the
moisture content of the blood. The blood will start to
mold very quickly and this mold will damage the evidence
item and the associated bloodstains. For damp or bloody
items one should use:
a) brown paper bags of appropriate size
b) earth guard bags
c) butcher paper that can be folded and
properly taped shut
 GENERAL GUIDE LINES FOR INVESTIGATION OFFICER:

Collect and Package Physical Evidence

2) Proper Labeling – Once items have been properly

packaged, they must be properly labeled. Proper labeling
includes the following:
a) The investigating agency’s case number
b) the scene site (especially if more than one scene is
involved)
c) a brief description of the evidence or the evidence
item number which correlates to an evidence log
d) the location where the evidence was obtained
within the scene
e) the collector’s name or badge no. and initials
f) the date and time collected
Evidence Collection

GENERAL GUIDE LINES FOR INVESTIGATION OFFICER:

Collect and Package Evidence

3) Chain of Custody – Once items have been properly
packaged and labeled, they must be transported to the
crime lab for further evaluation. A complete and correct
chain of custody must be maintained for all items. Often it
is not the evidence collector who transports the evidence
to the lab so a chain of custody log must be maintained
indicating the transfer of custody to and from every
individual who is involved in transporting or storing the
evidence until it gets to the crime lab. These include:
a) the collecting officer
b) the transportation officer
c) any evidence storage officer if the evidence is
stored prior to taking it to the lab
d) any further transportation officer
e) anyone who gets into the evidence for any reason
f) the laboratory evidence collection person(s)
Evidence Collection

GENERAL GUIDE LINES FOR INVESTIGATION OFFICER:

Beware of Contamination
Contamination occurs when DNA from another source
gets mixed in with the sample being collected.
 An investigator touches, sneezes, bleeds on a sample.
 Wear gloves and use disposable instruments
 Package items separately.
 Especially, do not mix known samples (from victim or
suspect) with unknown samples.
GENERAL GUIDE LINES FOR INVESTIGATION OFFICER:
Chain of Custody

 A record of individuals who have had

PHYSICAL possession of the evidence
 Critical in maintaining the integrity of
the evidence
 If DNA analysis results in a foreign
DNA type, it may be necessary to
identify persons who handled the
evidence
Evidence Collection
GENERAL GUIDE LINES FOR INVESTIGATION OFFICER:

Chain of Custody Components

 Identifiers that describe the evidence at

the time it was found
 Location
 Position
 Date/Time of Collection
 Packaging/Sealing information
Evidence Collection

Biological Evidence Collection :

Blood
Evidence Collection

DNA FINGER PRINTING KEY CONCEPT:

IDENTIFY PEOPLE AT MOLECULAR LEVEL

Evidence Collection

MEDICAL OFFICER’S JOB TO WITHDRAW BLOOD

Evidence Collection

LIQUID BLOOD AND FRESH

BLOOD STAINS

Blood that is in liquid pools should be picked up on a gauze

pad or other clean sterile cotton cloth and allowed to air dry
thoroughly, at room temperature.

It should be refrigerated or frozen as soon as possible and

brought to the Laboratory as quickly as possible. Delays
beyond 48 hours may make the samples useless.

If close to the Laboratory, deliver stained object immediately.

If unable to deliver to the Laboratory, or if the object must be

mailed, allow the stain to air dry completely before
packaging.
Evidence Collection

LIQUID BLOOD AND FRESH

BLOOD STAINS

Do not heat stained material or place it in bright

sunlight to dry. Hang clothing and similar articles in
a room where there is adequate ventilation.

If not completely dry, label and roll in paper or

place in a brown paper bag or box and seal and
label container. Place only one item in each
container.

Do not use plastic containers.

Evidence Collection

Liquids Blood

Liquids (generally standards)Blood

Refrigerate, do not freeze standards collected in
stoppered vacutainers.
Submit to the lab as soon as possible

STAINED CLOTHES:
On clothing, if possible, wrap the item
in clean paper, place the article in a
brown paper bag or box and seal and
label container. Do not attempt to
remove stains from the cloth.
LIQUID BLOOD VACUTAINERS COATED WITH EDTA
Evidence Collection

FTA CARD FOR BLOOD STAIN COLLECTIOON

1 Case detail marking

Provision

2.One Circle for one

specimen

Whatman™  Whatman International Ltd.

Evidence Collection

PLEASE DO NOT USE COTTON FOR LIQUID BODY FLUID

BLOOD STAIN COLLECTIOON

Filter Paper Clinical Gauze

Evidence Collection

BLOOD STAINED ITEMS NEED TO BE AIR DRIED AND PACKED SEPARATELY

Evidence Collection

Blood Stain on Weapon

Dry the knife. Don’t Remove stain

Blood Stain Evidence Collection
Blood Trails Away From Body
 Blood stains on wall/floor/hard surface and
other material
On small solid objects, send the whole stained object to the
Laboratory, after labeling and packaging.

On large solid objects, cover the stained area with clean paper
and seal the edges down with tape to prevent loss or
contamination. If impractical to deliver the whole object to the
Laboratory, scrape the stain onto a clean piece of paper, which
can be folded and placed in an envelope. Do not scrape directly
into evidence envelope. Scrape blood from objects using a
freshly washed and dried knife or similar tool. Wash and dry the
tool before each stain is scraped off. Seal and mark the
envelope.

Do not mix dried stains. Place each stain in a separate

envelope.

REMEMBER: Never attempt to wipe dried stains from an object

using a moistened cloth or paper.
HANDLING SEXUAL ASSAULT RELATED EXHIBITS
 "The ax forgets, the tree remembers."

Traumatized people
suffer damage to the
basic structures of
the self. They lose
trust in themselves,
in other people, in
God...The identity
they have formed
prior to the trauma is
irrevocably
destroyed.

Remember: A charge of rape is easy to make, hard to

prove, harder still Helping Victims Preserve DNA Evidence
to disprove
Procedure for rape victim examination:

 Police advise victim not to change clothes,

bathe before Medical Examination.
 Should not be examined without authority
of investigating PO/ magistrate.
 Consent of victim/ guardian, date/ time,
identified by escorting female PO.
Examine in POs presence.
 Examine without delay – vulva
tenderness, detection of spermatozoa.
Evidence Collection

Semen and Vaginal Secretion stains are the two

most commonly tested biological evidence.

Moisture allows for bacterial growth and

destroys DNA. Therefore:
 Foreign hair preserved and compared with
accused.
 Saliva / lipstick marks etc be carefully collected.
Evidence Collection

Forensic search of of Semen Stains

At the scene of a sexual assault, the

search for semen includes the corpse or
victim, underwear, bed sheets and
mattresses, carpeting, and flooring.

 Clothes are examined for the presence of blood and

seminal stains, tears and mud.
 Any marks or suspicious stains are preserved for
examination by the chemical examiner.
 Stands on clean sheet of paper, and anything that
falls is preserved – earth, buttons, hair.
 Foreign hair preserved and compared with accused.
Semen

1.Locate semen stain – often visible

 exhibit a stiff, crusty
appearance identify stain.
2.Possibly, blood stains may be
found type immediately
Remember: If not willfully removed from
the vaginal tract semen can be detected
up to 72 hours. It is advised to get patient
examined before 72 hrs.
Evidence Collection

Specimens to be collected

 Head hairs
 Pubic hair
 Blood group
 Urine drug screening
 Nail scrapings for blood or tissue
 Swabs - soiled areas of skin, anal,
rectum
 Swab - penis of accused.
Evidence Collection

Genital examination
 Examined by an O&G specialist.
 Genital organs examined for signs of
bleeding, bruises and lacerations.
 Matted pubic hair due to the presence of
semen is cut off for examination.
 Comb pubic hair for non-matching hair,
root study.
 Dried seminal stains on the external
genitals and thighs are scraped off by
means of a clean blunt knife for
subsequent examination.
Clothes

 Clothes are examined for the presence of

blood and seminal stains, tears and mud.
 Any marks or suspicious stains are
preserved for examination by the
chemical examiner.
 Stands on clean sheet of paper, and
anything that falls is preserved – earth,
buttons, hair.
 Foreign hair preserved to be compared
with accused.
 Evidence Collection

PAPER ENVELOPE

Air Drying on neat

paper
 Identification of Semen

Searching for Semen Stains

 May be on clothing,
skin, bedding, etc.
 Visual inspection;
yellowish-white
 Alternative light source:
look for fluorescence
 SEMEN SAMPLE SUBMISSION GUIDELINES

1. Air dry evidence before packing

2. Do not let two items of evidence come in contact with each
other
3. Package evidence in paper bags or envelopes
4. Package different pieces of evidence separately
Evidence Collection

VAGINAL SECRETION SLIDES:

Microscope

Vaginal Swab Smear

Note labeling on slide

Identification
Sperm Cellsof Semen
Under Microscope
Evidence Collection

Vaginal Swab Collection:

Vaginal Swab
Evidence Collection

SALIVA EXAMINATION:
 BUCCUL EPITHILIAL CELLS COLLECTION:

Saddam
Hussein’s
capture
verified with
DNA testing
conducted in
Rockville at
Armed Forces
DNA
Identification
Laboratory
 Blood
 Semen/
sperm
 Body tissue/
skin
 Bone/ bone
marrow
 Hair/ hair
roots
 Saliva/ bite
marks
 Urine
 Teeth/dental
 Fecal
material
 Mucous
Evidence Collection

Potential Sources of DNA Evidence

 Sources of Biological Evidence

 • Blood
 • Semen
 • Saliva
 • Urine
 • Hair
 • Teeth
 • Bone
 • Tissue
Evidence Collection

Skull
Human Bite Mark
Femur
 PACKING FOR BONE EXHIBITS
E
ENC
EVID

ENCE
EVID

EVIDENCE
Evidence Collection

Evidence Storage:

HAIR

 Foreign hair preserved and compared

with accused.
 DNA test raises doubts over Texas man's execution
            
AFP - Friday, November 12WASHINGTON (AFP) - – A Texas man was condemned to death and executed in 2000 on
the basis of hair that did not belong to him, according to the results of a recent DNA test.
A test by Mitotyping Technologies published by the Texas Observer magazine -- which fought a three-year legal battle to
gain access to the evidence -- showed that Claude Howard Jones was "excluded as the contributor of this questioned
hair."
Jones - who had a long criminal record - had insisted that he was waiting in the car when his accomplice killed Allen
Hilzendager during a liquor store robbery.
He was convicted of the 1989 murder, and denied several appeals, largely on the basis of that single strand of hair which
police found at the scene.
Forensic science was limited at the time to examining the hair under a microscope, where it appeared to belong to Jones.
The DNA test posted on the magazine's website found that the hair likely belonged instead to the victim.
Analyzing hair under a microscope was later abandoned after it was deemed inconclusive and obsolete with the
development of DNA testing.
Jones requested a DNA test and a stay of execution until it could be performed, but he was denied by then-governor
George W. Bush.
Documents obtained by the Texas Observer and the Innocence Project showed that "attorneys in the governor's office
failed to inform Bush that DNA evidence might exonerate Jones," the Observer wrote.
Bush, embroiled in the 2000 presidential election recount at the time of the execution, was a proponent of DNA testing in
death penalty cases and had previously halted another execution so key evidence could be tested.
"Because the DNA testing doesn't implicate another shooter, the results don't prove Jones' innocence," the Texas
Observer wrote.
"But the hair was the only piece of evidence that placed Jones at the crime scene. So while the results don't exonerate
him, they raise serious doubts about his guilt."
A Texas judge is currently considering whether another man executed on the basis of out-dated forensic evidence was
indeed innocent.
Cameron Todd Willingham was executed in 2004 for setting a 1991 fire that killed his three daughters. Experts have
testified that the evidence used to prove the fire was arson was flawed.
Evidence Collection

FORENSIC HAIR
EVIDENCE COLLECTION

HAIR SAMPLE STANDARDS: Whenever hair is collected the roots should be

included because considerable information can be obtained from the root
material.

HEAD OR SCALP HAIR The hair should be representative of the center,

front, back (including nape of the neck), and both sides of the scalp.
Approximately 50 head hairs should be collected. The sample should include
both pulled and combed hairs and include any variations in color and length.
If additional facial hairs are collected (i.e. sideburn or beard hairs), these
should be packaged separately.

PUBIC HAIR: When indicated by the circumstances, collect pubic hair.

Approximately 30 pubic hairs should be collected. The sample should be
collected and packaged in the same manner described above.
Collection and preservation of hair
evidence

• Have questioned and standard/reference

hairs come from same area of body and
adequate number of hair samples of each

• Forensic hair comparisons involve either

head hair or pubic hair

• Collection of 50 full-length hairs from all

areas of the scalp

• Minimum of 24 full-length hairs from pubic

area
Collection and preservation of hair
evidence continued

• Entire hair length collected because of

variance in morphological
characteristics
like color
• Pull hair out of skin or clipping at skin
line
• Hair samples collected from victim(s)
suspicious death during autopsy
Ascertaining present status forensic hair
examination

• DNA can individualize a human hair

• Nuclear DNA produce frequency of occurrences

as low as 1 in billions or trillions

• Mitochondrial DNA will not individualize human

hair but can exclude a significant portion of
population as hair contributors

• Mitochondrial DNA cannot distinguish

microscopically similar hairs from different
individuals who are maternally related
 COLLECTION, PRESERVATION AND MARKING OF HAIR
EVIDENCE

Before attempting specific procedures listed below, note the following

general precautions:
1. The size of the container should correspond to the size of the object.

2. Do not package wet evidence. Hair or objects containing hair evidence

should be air dried before being placed in sealed containers. Biological
stains degrade with time. This process is accelerated when items are wet
and sealed in airtight containers.

3 Do not package items on a surface without first thoroughly cleaning that

surface. Avoiding cross contamination between all evidence and
standards is imperative!

4. All seams of the packaging must be sealed to prevent the loss of trace
evidence.

5. Label all evidence containers with submitter’s initials, ID/badge number,

agency name, case number, item number, source, and date.
 HAIR AS DNA EVIDENCE

Fallen Out Cut Torn Pulled Out Pulled Out

EVIDENCE PROPERTY BAG

EVIDENCE
Case No.
Police Station
 EVID
ENCE
Case
N
Police o.
Statio
n
EVIDENCE PROPERTY BAG
 HAIR AS DNA EVIDENCE

CE
EN
ID
EV

Self sealing plastic bag

 ITS
CRIME EXHIB
To:
The Director
CFSL,
ad
30-Gorachand Ro
Kolkata-14

Case
No.123
P.S.:XYZ
 COME TO CFSL WITH ALL THESE EXHIBITS AND DOCUMENTS:

Item
No. Check list

1 Brief history of the case. Yes/No

2 Duly sealed and marked exhibits and controls. Yes/No
3 List of exhibits/controls forwarded with details of collection and seals Yes/No
4 Duly filled "Biological Sample Donor Card", in duplicate for control samples. Yes/No
(DNA Cases only)
5 Relevant questions for examination asked Yes/No
6 Sample seal(s) in sealing wax on parcels/packets intact and legible. Yes/No
7 Copy of Post- mortem or medico- legal report of victim(s) & suspect(s), if Yes/No
conducted.
8 Letter of authority in favour of Director, Forensic Science Laboratory “to Yes/No
examine the exhibits, and if necessary, to take them to places or remove
portions for the purpose of examination”.

9 Case forwarded by an officer not below the rank of Superintendent of Yes/No

Police/Deputy Commissiner of Police (SP/DCP) or officer of equivalent rank
from Military, Custom and other agencies.
Thanks