A PAPER PRESENTATION

ON

DNA FINER PRINTING

A NEW INNOVATION IN FORENSIC SCIENCE
OR
CRIME INVESTIGATION

ABSTRACT
DNA Fingerprinting, method of identification that compares fragments of
deoxyribonucleic acid (DNA) It is sometimes called DNA typing. DNA is the genetic
material found within the cell nuclei of all living things. In mammals the strands of
DNA are grouped into structures called chromosomes. With the exception of
identical twins, the complete DNA of each individual is unique.

A DNA fingerprint is constructed by first extracting a DNA sample from body

tissue or fluid such as hair, blood, or saliva. The sample is then segmented using
enzymes, and the segments are arranged by size using a process called
electrophoresis. The segments are marked with probes and exposed on X-ray film,
where they form a characteristic pattern of black bars—the DNA fingerprint. If the
DNA fingerprints produced from two different samples match, the two samples
probably came from the same person.

DNA fingerprinting was first developed as an identification technique in 1985.

Originally used to detect the presence of genetic diseases, DNA fingerprinting
soon came to be used in criminal investigations and forensic science. The first
criminal conviction based on DNA evidence in the United States occurred in 1988.
In criminal investigations, DNA fingerprints derived from evidence collected at the
crime scene are compared to the DNA fingerprints of suspects. The DNA evidence
can implicate or exonerate a suspect.

Generally, courts have accepted the reliability of DNA testing and admitted DNA
test results into evidence. However, DNA fingerprinting is controversial in a
number of areas: the accuracy of the results, the cost of testing, and the possible
misuse of the technique.

A DNA fingerprint is made by analyzing the sizes of DNA fragments produced from
a number of different sites in the genome that vary in length.

The more common the length variation at a particular site and the greater the number
the sites analyzed, the more informative the fingerprint.

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In this paper we shall discuss about how DNA finger printing is being done, what are
its applications and the uses of DNA finger printing

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 DNA FINGER PRINTING BASICS

Different individuals carry different alleles.

Most alleles useful for DNA fingerprinting differ on the basis of the number of
repetitive DNA sequences they contain.
A site with three alleles useful for DNA finger printing

DNA fragments of different size will be produced by a restriction enzyme that cuts at
the points shown by the arrows.
HOW IS DNA FINGER PRINTING DONE?
1. Performing a Southern Blot
2. VNTRs
SOUTHERN BLOT:

VNTR’S:

Every strand of DNA has pieces that contain genetic information which informs an
organism's development (exons) and pieces that, apparently, supply no relevant
genetic information at all (introns). Although the introns may seem useless, it has
been found that they contain repeated sequences of base pairs. These sequences,
called Variable Number Tandem Repeats (VNTRs), can contain anywhere from
twenty to one hundred base pairs.

Because VNTR patterns are inherited genetically, a given person's VNTR pattern is
more or less unique. The more VNTR probes used to analyze a person's VNTR
pattern, the more distinctive and individualized that pattern, or DNA fingerprint,will
be.

→THE DNA FRAGMENTS ARE SEPERATED ON THE BASIS OF THE SIZE:

The technique is gel electrophoresis.

 A DNA FINGER PRINT

When many genes are analyzed, each with many different alleles, the chance that two
patterns match by coincidence is vanishingly small.
 MECHANICS OF DNA FINGER PRINT

The nucleus of every cell in the human body contains deoxyribonucleic acid or DNA,
a biochemical molecule that is made up of nearly three-billion nucleotides. DNA
consists of four different nucleotides, adenine (A), thymine (T), guanine (G), and
cytosine (C), which are strung together in a sequence that is unique to every
individual. The sequence of A, T, G, and C in human DNA can be found in more
combinations or variations than there are humans. The technology of DNA
fingerprinting is based on the assumption that no two people have the same DNA
sequence.

The DNA from a small sample of human tissue can be extracted using biochemical
techniques. Then the DNA can be digested using a series of enzymes known as
restriction enzymes, or restriction endonucleases. These molecules can be thought of
as chemical scissors, which cut the DNA into pieces. Different endonucleases cut
DNA at different parts of the nucleotide sequence. For example, the endonuclease
called SmaI cuts the sequence of nucleotides CCCGGG between the third cytosine
(C) and the first guanine (G).

After being exposed to a group of different restriction enzymes, the digested DNA
undergoes gel electrophoresis. In this biochemical analysis technique, test samples of
digested DNA are placed in individual lanes on a sheet of an agarose gel that is made
from seaweed. A separate lane contains control samples of DNA of known lengths.
The loaded gel is then placed in a liquid bath and an electric current is passed through
the system. The various fragments of DNA are of different sizes and different
electrical charges. The pieces move according to their size and charge with the
smaller and more polar ones travelling faster. As a result, the fragments migrate down
the gel at different rates.

After a given amount of time, the electrical current in the gel electrophoresis
instrumentation is shut off. The gel is removed from the bath and the DNA is blotted
onto a piece of nitrocellulose paper. The DNA is then visualized by the application of
radioactive probe that can be picked up on a piece of x-ray film. The result is a film
that contains a series of lines showing where the fragments of DNA have migrated.
Fragments of the same size in different lanes indicate the DNA has been broken into
segments of the same size. This demonstrates a similarity between the sequences
under test.

Different enzymes produce different banding patterns and normally several different
endonucleases are used in conjunction to produce a high definition banding pattern on
the gel. The greater the number of enzymes used in the digestion, the finer the
resultant resolution.

In DNA fingerprinting, scientists focus on segments of DNA in which nucleotide

sequences vary a great deal from one individual to another. For example, five to ten
percent of the DNA molecule contains regions that repeat the same nucleotide
sequence many times, although the number of repeats varies from person to person.
Jeffreys targeted these long repeats called variable number of tandem repeats
(VNTRs) when he first developed DNA fingerprinting. The DNA of each person also
has different restriction fragment sizes, called restriction fragment length
polymorphisms (RFLPs), which can be used as markers of differences in DNA
sequences between people. Today, technicians also use short tandem repeats (STRs)
for DNA fingerprinting. STRs are analyzed using polymerase chain reaction or PCR,
a technique for mass-producing sequences of DNA. PCR allows scientists to work
with degraded DNA.

PRACTICAL APPLICATIONS OF DNA FINGER PRINTING

1. Paternity and Maternity

Because a person inherits his or her VNTRs from his or her parents, VNTR patterns
can be used to establish paternity and maternity. The patterns are so specific that a
parental VNTR pattern can be reconstructed even if only the children's VNTR
patterns are known (the more children produced, the more reliable the reconstruction).
Parent-child VNTR pattern analysis has been used to solve standard father-
identification cases as well as more complicated cases of confirming legal nationality
and, in instances of adoption, biological parenthood.

2. Criminal Identification and Forensics

DNA isolated from blood, hair, skin cells, or other genetic evidence left at the scene
of a crime can be compared, through VNTR patterns, with the DNA of a criminal
suspect to determine guilt or innocence. VNTR patterns are also useful in establishing
the identity of a homicide victim, either from DNA found as evidence or from the
body itself.

3. Personal Identification
The notion of using DNA fingerprints as a sort of genetic bar code to identify
individuals has been discussed, but this is not likely to happen anytime in the
foreseeable future. The technology required to isolate, keep on file, and then analyze
millions of very specified VNTR patterns is both expensive and impractical. Social
security numbers, picture ID, and other more mundane methods are much more likely
to remain the prevalent ways to establish personal identification.

A DNA fingerprint used in a murder case

The defendant stated that the blood on his clothing was his
What are we looking at? How was it produced?

Use as a forensic tool. DNA fingerprinting is now an important tool in the arsenal of
forensic chemists. It is used in forensics to examine DNA samples taken from a crime
scene and compare them to those of a suspect. Criminals almost always leave
evidence of their identity that contains DNA at the crime scene—hair, blood, semen,
or saliva. These materials can be carefully collected from the crime scene and
fingerprinted

Although DNA fingerprinting is scientifically sound, the use of DNA fingerprinting

in courtrooms remains controversial. There are several objections to its use. Lawyers
who misrepresent the results of DNA fingerprints may confuse jurors. DNA
fingerprinting relies on the probability that individuals will not produce the same
banding pattern on a gel after their DNA has been fingerprinted. Establishing this
probability relies on population statistics. Each digested fragment of DNA is given a
probability value. The value is determined by a formula relating the combination of
sequences occurring in the population. There is concern that not enough is known
about the distribution of banding patterns of DNA in the population to express this
formula correctly. Concerns also exist regarding the data collection and laboratory
procedure associated with DNA fingerprinting procedures. For example, it is possible
that cells from a laboratory technician could be inadvertently amplified and run on the
gel. However, because each person has a unique DNA sequence and this sequence
cannot be altered by surgery or physical manipulation, DNA fingerprinting is an
important tool for solving criminal cases

SLT 3/8/05

Some applications of DNA finger printing in justice system.

In addition to the advantages there are also some problems with DNA finger printing.
They are:

1.GENERATING A HIGH PROBABILITY.

The probability of a DNA fingerprint belonging to a specific person needs to be

reasonably high--especially in criminal cases, where the association helps establish a
suspect's guilt or innocence. Using certain rare VNTRs or combinations of VNTRs to
create the VNTR pattern increases the probability that the two DNA samples do
indeed match (as opposed to look alike, but not actually come from the same person)
or correlate (in the case of parents and children).

2. PROBLEMS WITH DETERMINING PROBABILITY

A. POPULATION GENETICS

B.TECHNICAL DIFFICUTLTIES.

HENCE DNA PRINTING IS USED AS A TOOL IN CRIME INVESTIGATION AND

IS HELPING THE OCIETY IN IDENTIFYING THE CRIMES.ALSO THE
PROBLEMS ARE BEING OVER COMED BY THE GENETIC ENGINEERING
PROCESSES.

THANK YOU.