What is a DNA fingerprint?

DNA fingerprinting is a laboratory technique used to

determine the probable identity of a person based on the
nucleotide sequences of certain regions of human DNA that
are unique to individuals. DNA fingerprinting is used in a
variety of situations, such as criminal investigations, other
forensic purposes and paternity testing. In these situations,
one aims to “match” two DNA fingerprints with one another,
such as a DNA sample from a known person and one from an
unknown person.

Background
 Almost every cell? in our body contains our DNA?.
 On average, about 99.9 per cent of the DNA between
two humans is the same.
 The remaining percentage is what makes us unique
(unless you are an identical twin!).
 Although this might sound like a small amount, it means
that there are around three million base pairs? that are
different between two people. These differences can be
compared and used to help distinguish you from
someone else.
 Minisatellites are short sequences (10-60 base pairs
long) of repetitive DNA that show
greater variation? from one person to the next than
other parts of the genome?. This variation is exhibited in
the number of repeated units or ‘stutters’ in the
minisatellite sequence.
  The first minisatellite was discovered in 1980.

DNA fingerprinting
 DNA fingerprinting was invented in 1984 by Professor Sir
Alec Jeffreys after he realised you could detect
variations in human DNA, in the form of these
minisatellites.
 DNA fingerprinting is a technique that simultaneously
detects lots of minisatellites in the genome to produce a
pattern unique to an individual. This is a DNA
fingerprint.
 The probability of having two people with the same DNA
fingerprint that are not identical twins is very small.
 Just like your actual fingerprint, your DNA fingerprint is
something you are born with, it is unique to you.

How was the first DNA fingerprint produced?

1.The first step of DNA fingerprinting was to extract DNA
from a sample of human material, usually blood.
2.Molecular ‘scissors’, called restriction enzymes?, were
used to cut the DNA. This resulted in thousands of
pieces of DNA with a variety of different lengths.
3.These pieces of DNA were then separated according to
size by a process called gel electrophoresis?:
 The DNA was loaded into wells at one end of a

porous gel, which acted a bit like a sieve.

 An electric current was applied which pulled the

negatively-charged DNA through the gel.

 The shorter pieces of DNA moved through the gel

easiest and therefore fastest. It is more difficult for

 the longer pieces of DNA to move through the gel
so they travelled slower.
 As a result, by the time the electric current was

switched off, the DNA pieces had been separated

in order of size. The smallest DNA molecules were
furthest away from where the original sample was
loaded on to the gel.
4.Once the DNA had been sorted, the pieces of DNA were
transferred or ‘blotted’ out of the fragile gel on to a
robust piece of nylon membrane and then ‘unzipped’ to
produce single strands of DNA.
5.Next the nylon membrane was incubated with
radioactive probes.
 Probes are small fragments of minisatellite DNA

tagged with radioactive phosphorous.

 The probes only attach to the pieces of DNA that

they are complementary? to – in this case they

attach to the minisatellites in the genome.
6.The minisatellites that the probes have attached to were
then visualised by exposing the nylon membrane to X-
ray film.
 When exposed to radioactivity a pattern of more

than 30 dark bands appeared on the film where

the labelled DNA was. This pattern was the DNA
fingerprint.
 To compare two or more different DNA fingerprints

the different DNA samples were run side-by-side

on the same electrophoresis gel.
 Illustration showing the steps in DNA fingerprinting.
Image credit: Genome Research Limited

DNA profiling
 Modern-day DNA profiling is also called STR analysis and
relies on microsatellites rather than the minisatellites
used in DNA fingerprinting.
 Microsatellites, or short tandem repeats (STRs), are the
shorter relatives of minisatellites usually two to five
base pairs long. Like minisatellites they are repeated
many times throughout the human genome, for
example ‘TATATATATATA’.

DNA Fingerprinting Steps

Alec Jeffreys developed this technique in which he used satellite DNAs also
called VNTRs (Variable Number of Tandem Repeats) as a probe because it showed the high
level of polymorphism.

Following are the steps involved in DNA fingerprinting:

Isolating the DNA.

Digesting the DNA with the help of restriction endonuclease enzymes.

Separating the digested fragments as per the fragment size by the process of electrophoresis.

Blotting the separated fragments onto synthetic membranes like nylon.

Hybridising the fragments using labelled VNTR probes.

 ↓

Analysing the hybrid fragments using autoradiography.

How is a DNA profile produced today?

1.DNA is extracted from a biological sample. STR analysis
is incredibly sensitive so it only needs a tiny amount of
someone’s DNA to produce an accurate result. As a
result the DNA can be extracted from a wider range of
biological samples, including blood, saliva and hair.
2.Unlike the original DNA fingerprinting method, DNA
profiling does not use restriction enzymes to cut the
DNA. Instead it uses the polymerase chain reaction
(PCR)? to produce many copies of specific STR
sequences.
 PCR is an automated procedure that generates lots

of copies of a specific sequence of DNA. It only

requires small amounts of DNA to start with and
can even make copies from a DNA sample that is
partially degraded.
?
 In PCR small bits of DNA called primers bind to

complementary sequences of the DNA of interest

and mark the starting point for the copying of the
DNA of interest.
 In STR analysis the primers used in the PCR are

designed to attach to either end of the STR

sequence of interest.
 The primers for each STR is labelled with a specific

coloured fluorescent tag. This makes it easier to

identify and record the STR sequences after PCR.
3.Once enough copies of the sequence have been
produced by PCR, electrophoresis is used to separate
the fragments according to size.
4.Each fragment passes by a laser which causes the
fragments with fluorescent tags to glow with a specific
colour. The output is displayed as a series of coloured
peaks (as shown in the image below) highlighting the
colour and length of each STR sequence.
 Illustration showing the steps in DNA profiling.
Image credit: Genome Research Limited

 The more STR sequences that are tested, the more

accurate the test is at identifying someone.
 Other STRs used for forensic purposes are called Y-STRs,
which are derived solely from the male Y chromosome?.
This is useful for identifying a male perpetrator from
mixed DNA samples.
 Only one person in every 10 million million
(10,000,000,000,000) will have a particular STR profile.
With the world human population estimated at only
7,100 million (7,100,000,000) it is therefore extremely
unlikely you will share the same profile as someone
else, unless you are an identical twin.

Solving crime
 DNA profiles are very useful in forensics because only a
tiny sample of human material left behind after a crime
may be sufficient to identify someone.
 In the UK, a complete DNA profile consists of 11 STR
sequences plus a sex determiner to confirm if the
profile is from a man or a woman. Now all new profiles
include an additional five STR sequences to provide
consistency across borders in Europe.
 In the USA, the Federal Bureau of Investigation (FBI)
recommends that 13 STR sequences are tested. Many
states are increasing the number of STR sequences
tested to enable more efficient investigations across
state borders.
 A match made between a crime scene profile and an
individual profile identifies a possible suspect.
 A match made between different crime scene profiles
indicates a repeat offender at work.
 The police may use this DNA evidence to support other
evidence to help prosecute someone for a crime.
Complete DNA profiles give very reliable matches and
may provide strong evidence that a suspect is guilty or
innocent of a crime.
Illustration showing a comparison of a DNA fingerprint from
a crime scene and DNA fingerprints from two suspects. The
DNA fingerprint from suspect 2 matches that taken from the
crime scene.
Image credit: Genome Research Limited
How are DNA profiles stored?
 The UK was the first country to set up a national
database of DNA profiles in 1995.
 The UK National DNA Database holds the DNA profiles
from a select number of UK individuals, most of which
are linked to serious crimes.
 The Protection of Freedom Act 2013 ensured that
1,766,000 DNA profiles taken from innocent adults and
children were deleted from the UK National DNA
Database.
 Most countries now have a national DNA database.

Linking blood relatives

 You get half of your DNA from your mother and half
from your father. STRs are therefore passed down from
parents to their children.
 DNA profiling can be used to help confirm whether two
people are related to one another and is commonly
used to provide evidence that someone is, or is not, the
biological parent of a child.
 DNA profiling can also be used to identify victims of
crime or major disasters and help bring separated
families back together.
 DNA profiling has a high success rate and very low false-
positive rate.
HISTORY OF DNA FINGERPRINTING USED IN LAW
AND CRIME

Forensic Science and DNA evidence

DNA fingerprinting was first used in forensic science in 1986
when police in the UK requested Dr. Alec J. Jeffreys, of
University of Leicester, to verify a suspect’s confession that
he was responsible for two rape-murders. Tests proved that
the suspect had not committed the crimes.
The first person to be convicted on the basis of DNA evidence
in the UK was Robert Melias in 1987 (9,10). In the same year
in the US, Tommy Lee Andrews was convicted in a rape case
based on DNA evidence (9), in which his DNA profile was
matched with that of semen traces recovered from the
victim. Two other important early cases gave much impetus
to the use of DNA evidence: They were, the case of Glen Dale
Woodal versus the State of West Virginia in 1992 and the
multiple murder trial of Timothy Wilson Spencer versus the
state of Virginia in 1994. The DNA evidence in the Woodal
case exonerated him while that of the Spencer case resulted
in his conviction and sentencing to the death penalty.

ADMISSIBILITY IN COURT

Admissibility of DNA evidence was seriously challenged for

the first time in a case in the New York Supreme Court in
1989. Jose Castro was accused of murdering one Vimla Pence
and her two year old daughter. Although a blood stain on
Castro’s watch was matched to the victim, this evidence per
se was not instrumental to his conviction. He was convicted
after admitting to the crime. In this case, the DNA tests
conducted by Life Code Corporation did not include a specific
test for human blood and also did not include blind testing
protocols in the attempt to link the stain to the victims.
Furthermore, the laboratory in the above case had used
contaminated probes and did not provide the worksheets
and other manuscripts relating to the testing. Hence the
court issued many directive guidelines regarding the test
procedures and maintenance of laboratory results and
reports as well as explanations for probability calculations
and recording of observed defects or laboratory errors. The
need to identify and document chain of custody and allowing
access to data, methodology and actual results for an
independent expert to review were also instructed.

In another case in 1989, the Supreme Court of Minnesota

had also refused to admit the DNA evidence analyzed by a
private forensic laboratory. The court noted that the
laboratory did not comply with appropriate standards and
controls. In particular the court castigated the laboratory for
failure to reveal its underlying population data and testing
methods. Such secrecy precluded replication of the test.

Thus, courts have denounced improper application of DNA

scientific techniques to particular cases, especially when used
to declare matches based on frequency estimates. However,
DNA testing when properly applied is generally accepted as
admissible and currently in many countries, DNA evidence is
routinely used as evidence. As stated in the US National
Research Council’s (NRC) 1996 report (10) on DNA
evidence, “The state of the profiling technology and the
methods for estimating frequencies and related statistics
have progressed to the point where the admissibility of
properly collected and analyzed DNA data should not be in
doubt”

The FBI measures the success of the CODIS program by

counting the crimes it helps to solve. The “cold hit” (20), is
defined as a match which provides the police with an
investigative lead that would not otherwise have been
developed. The following two cases illustrate typical CODIS
hits.

Case 1:

St. Paul, Minnesota, November 1994: A man wearing a nylon

stocking over his face and armed with a knife jumped out
from behind bushes and forced a woman who was walking by
to perform oral sex. Semen recovered from the victim’s skirt
and saliva was analyzed using DNA technology. The resulting
profile was searched against Minnesota’s CODIS database.
The search identified Terry Lee Anderson, who confessed and
he is now in prison.

Case 2:

Tallahassee Florida, February 1995: The Florida Department

of Law Enforcement linked semen found on a Jane Doe -
rape-homicide victim to a convicted offender’s DNA profile.
The suspect’s DNA was collected, analyzed, and stored in a
CODIS database. A match was later identified to be of a
convicted rapist was timely; it prevented the offender’s
release on parole scheduled eight days later.

The organization and structure of CODIS can be a model for

establishing such a system in Malaysia. The Malaysian
parliament has recently passed an Act to collect and type
DNA from convicted offenders hence paving the way for such
a system to be implemented.
DNA Fingerprinting Applications
As discussed earlier the technique of fingerprinting is used for DNA analysis in forensic tests
and paternity tests. Apart from these two fields, it is also used in determining the frequency of
a particular gene in a population which gives rise to diversity. In case of the change in gene
frequency or genetic drift, Fingerprinting can be used to trace the role of this change in
evolution.
BIBLIOGRAPHY
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3561883/
#:~:text=DNA%20fingerprinting%20was%20first%20used,had
%20not%20committed%20the%20crimes.
https://www.genome.gov/genetics-glossary/DNA-
Fingerprinting
https://byjus.com/biology/dna-fingerprinting/
https://www.yourgenome.org/facts/what-is-a-dna-
fingerprint/