DNA Profiling

(DNA fingerprinting) A method of Forensic identification

What is DNA Profiling?

A technique used by scientists to distinguish between individuals of the same species using only samples of their DNA

Who Invented it?

The process of DNA fingerprinting was invented by Alec Jeffreys at the University of Leicester in 1985. He was knighted in 1994.

 The

primary method of assessing similarities is by use of DNA fingerprinting or DNA restriction analysis. This process makes use of special proteins called restriction enzymes and sections of the chromosome called tandem repeats

How do you figure out that someones DNA is more similar to anothers?

Tandem Repeats
A

region of the chromosome that contains multiple copies of a core DNA sequence that are arranged in a repeating fashion Repeats act as fillers or spacers between coded sections of DNA All humans have the same type of repeats but there is tremendous variation in the number of repeats that each of us has.

Stages of DNA Profiling

Stage 1: Cells are broken down to release DNA

If only a small amount of DNA is available it can be amplified using the polymerase chain reaction (PCR)

Stages of DNA Profiling

Step 2: The DNA is cut into fragments using restriction enzymes.

Each restriction enzyme cuts DNA at a specific base sequence.

 SSR=

Simple sequence Repeats

or

SSLP=

Simple sequence length polymorphism

Simple-Sequence Length Polymorphisms (SSLPs)

D
VNTRs :
Variation in the Number of Tandem Repeats or Mini-satellite Molecular Markers

d
Probe binds repetitive sequences
Restriction target sites are outside the repetitive array. The basic unit of the array is indicated by the arrows.

The number of repeated units in a tandem array is variable. Individuals heterozygous for different numbers of tandem repeats can be detected, and the heterozygous site (s) used as a marker (s) for mapping.

This VNTR locus will form two bands on a Southern blot: one long and one short. Similar to an RFLP locus, this heterozygous site can be used for genetic mapping. At present, VNTR analysis is rapidly performed using PCR.

Restriction Fragment Length Polymorphisms (RFLP)

RFLPs

are different fragment lengths of base pairs that result from cutting a DNA molecule with a restriction enzyme It is the length differences associated with DNA strands or RFLPs that allow one to distinguish one person from another.

Stages of DNA Profiling

The

sections of DNA that are cut out are called restriction fragments. yields thousands of restriction fragments of all different sizes because the base sequences being cut may be far apart (long fragment) or close together (short fragment).

This

Stages of DNA Profiling

Stage 3: Fragments are separated on the basis of size using a process called gel electrophoresis. DNA fragments are injected into wells and an electric current is applied along the gel.

Stages of DNA Profiling

DNA is negatively charged so it is attracted to the positive end of the gel. The shorter DNA fragments move faster than the longer fragments. DNA is separated on basis of size.

Stages of DNA Profiling

A radioactive material is added which combines with the DNA fragments to produce a fluorescent image. A photographic copy of the DNA bands is obtained.

Stages of DNA Profiling

Stage 4: The pattern of fragment distribution is then analysed.

Uses of DNA Profiling

DNA

profiling is used to solve crimes and medical problems

Crime
Forensic

science is the use of scientific knowledge in legal situations. The DNA profile of each individual is highly specific. The chances of two people having exactly the same DNA profile is 30,000 million to 1 (except for identical twins).

Biological materials used for DNA profiling

Blood Hair Saliva Semen Body tissue cells DNA samples have been obtained from vaginal cells transferred to the outside of a condom during sexual intercourse.

DNA Profiling can solve crimes

The pattern of the DNA profile is then compared with those of the victim and the suspect. If the profile matches the suspect it provides strong evidence that the suspect was present at the crime scene (NB:it does not prove they committed the crime). If the profile doesnt match the suspect then that suspect may be eliminated from the enquiry.

Example
A

violent murder occurred. The forensics team retrieved a blood sample from the crime scene. They prepared DNA profiles of the blood sample, the victim and a suspect as follows:

Was the suspect at the crime scene?

Suspects Profile Blood sample from crime scene Victims profile

Solving Medical Problems

DNA profiles can be used to determine whether a particular person is the parent of a child. A childs paternity (father) and maternity(mother) can be determined. This information can be used in Paternity suits Inheritance cases Immigration cases

Example: A Paternity Test

By

comparing the DNA profile of a mother and her child it is possible to identify DNA fragments in the child which are absent from the mother and must therefore have been inherited from the biological father.

Is this man the father of the child?

Mother Child Man

An Example Using EcoR I for a Question of Paternity

Recall: EcoR I cuts only at GAATTC

EcoR I cuts a similar section of DNA on Bob, Larry, and Mary After the cut how many fragments Bob, Larry, and Mary have? Answer: 2, 3, and none

Example Cont
After the DNA is cut with EcoR I, Bobs, Larrys and Marys fragments are placed in different lanes on an agarose gel The fragments are then subjected to an electric field The smaller fragments move faster, the larger ones move slower This process of separating the fragments by length is called electrophoresis.

Resulting Picture after Electrophoresis

The

bigger fragments are near the top

Paternity Test

In

general the childs DNA must be a combination of Marys DNA and one of the men. Which man is the father? Answer: Larry

Famous cases

In 2002 Elizabeth Hurley used DNA profiling to prove that Steve Bing was the father of her child Damien

Famous Cases
Colin Pitchfork was the first criminal caught based on DNA fingerprinting evidence. He was arrested in 1986 for the rape and murder of two girls and was sentenced in 1988.

Famous Cases

O.J. Simpson was cleared of a double murder charge in 1994 which relied heavily on DNA evidence. This case highlighted lab difficulties.

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