DNA fingerprinting, in genetics, method of isolating and identifying

variable elements within the base-pair sequence of DNA (deoxyribonucleic

acid). The technique was developed in 1984 by British geneticist Alec
Jeffreys, after he noticed that certain sequences of highly variable DNA
(known as minisatellites), which do not contribute to the functions of genes,
are repeated within genes. Jeffreys recognized that each individual has
a unique pattern of minisatellites (the only exceptions being multiple
individuals from a single zygote, such as identical twins).

Learn how a genetic fingerprint is made using agarose gel, Southern blotting, and a
radioactive DNA probeLearn about how DNA is extracted, treated with restriction
enzymes, and sequenced using gel electrophoresis to create a genetic fingerprint.
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The procedure for creating a DNA fingerprint consists of first obtaining a
sample of cells, such as skin, hair, or blood cells, which contain DNA. The
DNA is extracted from the cells and purified. In Jeffreys’s original approach,
which was based on restriction fragment length polymorphism (RFLP)
technology, the DNA was then cut at specific points along the strand
with proteins known as restriction enzymes. The enzymes produced
fragments of varying lengths that were sorted by placing them on a gel and
then subjecting the gel to an electric current (electrophoresis): the shorter
the fragment, the more quickly it moved toward the positive pole (anode).
The sorted double-stranded DNA fragments were then subjected to a
blotting technique in which they were split into single strands and
transferred to a nylon sheet. The fragments underwent autoradiography in
which they were exposed to DNA probes—pieces of synthetic DNA that
were made radioactive and that bound to the minisatellites. A piece of X-
ray film was then exposed to the fragments, and a dark mark was produced
at any point where a radioactive probe had become attached. The resultant
pattern of marks could then be analyzed.

The assay developed by Jeffreys has been supplanted by approaches that

are based on the use of the polymerase chain reaction (PCR) and so-called
microsatellites (or short tandem repeats, STRs), which have shorter repeat
units (typically 2 to 4 base pairs in length) than minisatellites (10 to more
than 100 base pairs in length). PCR amplifies the desired fragment of DNA
(e.g., a specific STR) many times over, creating thousands of copies of the
fragment. It is an automated procedure that requires only small amounts of
DNA as starting material and works even with partially degraded DNA.
Once an adequate amount of DNA has been produced with PCR, the exact
sequence of nucleotide pairs in a segment of DNA can be determined by
using one of several biomolecular sequencing methods. Automated
equipment has greatly increased the speed of DNA sequencing and has
made available many new practical applications, including pinpointing
segments of genes that cause genetic diseases, mapping the human
genome, engineering drought-resistant plants, and producing
biological drugs from genetically altered bacteria.

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An early use of DNA fingerprinting was in legal disputes, notably to help

solve crimes and to determine paternity. Since its development, DNA
fingerprinting has led to the conviction of numerous criminals and to the
freeing from prison of many individuals who were wrongly convicted.
However, making scientific identification coincide exactly with legal proof is
often problematic. Even a single suggestion of the possibility of error is
sometimes enough to persuade a jury not to convict a suspect. Sample
contamination, faulty preparation procedures, and mistakes in
interpretation of results are major sources of error. In addition, RFLP
requires large amounts of high-quality DNA, which limits its application
in forensics. Forensic DNA samples frequently are degraded or are
collected postmortem, which means that they are lower-quality and subject
to producing less-reliable results than samples that are obtained from a
living individual. Some of the concerns with DNA fingerprinting, and
specifically the use of RFLP, subsided with the development of PCR- and
STR-based approaches.
 i. .

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.
DNA fingerprinting. I think a lot of people are first introduced to DNA
fingerprinting while watching crime shows. An officer collects some samples
from the crime scene. They put it in a tube. And then an hour later, they hold
up a brightly colored gel, squint at it, and say, aha, we have a match for the
killer's DNA. Then the show is over. Of course, that isn't exactly how things
work in real life. But DNA fingerprinting is an important part of forensic
science. Although it can't really tell you exactly who committed a crime, it can
be used to help narrow down a list of suspects based on how well their DNA
matches the samples that were found at the crime scene. Investigators can
also use the DNA results to search specific databases to find other potential
suspects.
What is DNA Fingerprinting?
Satellite DNA regions are stretches of repetitive DNA which do not code for any specific
protein. These non-coding sequences form a major chunk of the DNA profile of humans.
They depict a high level of polymorphism and are the basis of DNA fingerprinting. These
genes show a high level of polymorphism in all kind of tissues as a result of which they
prove to be very useful in forensic studies.

Any piece of DNA sample found at a crime scene can be analysed for the level of
polymorphism in the non-coding repetitive sequences. After the DNA profile is traced, it
becomes easier to find the criminal by performing the DNA fingerprinting for the
suspects.

Apart from crime scenes, Fingerprinting applications also prove useful in finding the
parents of an unclaimed baby by conducting a paternity test on a DNA sample from the
baby.

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

Discovery, development, and current

applications of DNA identity testing

Genetic identity testing involves identifying the patterns of genetic material

that are unique to almost every individual. Although over 99% of the DNA
sequences in the human genome are identical between individuals, a small
number of sequence differences are used to distinguish all humans (1). Those
different sequences are usually targeted for identity testing. The techniques
that are applied in identity testing are DNA fingerprinting, DNA profiling, and
DNA typing. Although there are some technical differences between these
tests, the terms have been used interchangeably.

DNA Fingerprinting is also called Restriction Fragment

Length Polymorphisms (RFLPs) Analysis

Restriction enzyme recognition sites are short (only a few nucleotides long),
sequence-specific palindromes, and may be found throughout the genome.
Thus, differences in DNA sequences in the genomes of individuals will lead to
differences in distribution of restriction enzyme recognition sites that can be
visualized as distinct fingerprints using a technique called gel electrophoresis
(see the seciton on Gel Electrophoresis below). Restriction fragment
length polymorphism (RFLP) analysis compares DNA banding patterns of
different DNA samples after restriction digestion.

RFLP analysis has many practical applications in both medicine and forensic
science. For example, epidemiologists use RFLP analysis to track and identify
the source of specific microorganisms implicated in outbreaks of food
poisoning or certain infectious diseases. RFLP analysis can also be used on
human DNA to determine inheritance patterns of chromosomes with variant
genes, including those associated with heritable diseases or to establish
paternity.
Forensic scientists use RFLP analysis as a form of DNA fingerprinting, which
is useful for analyzing DNA obtained from crime scenes, suspects, and
victims. DNA samples are collected, the numbers of copies of the sample
DNA molecules are increased using PCR, and then subjected to restriction
enzyme digestion and agarose gel electrophoresis to generate specific
banding patterns. By comparing the banding patterns of samples collected
from the crime scene against those collected from suspects or victims,
investigators can definitively determine whether DNA evidence collected at
the scene was left behind by suspects or victims.

Figure 2: RFLP analysis can be used to differentiate DNA sequences. In this example, a normal
chromosome is digested into two fragments, whereas digestion of a mutated chromosome
produces only one fragment. The small red arrows pointing to the two different chromosome
segments show the locations of the restriction enzyme recognition sites. After digestion and
agarose gel electrophoresis, the banding patterns reflect the change by showing the loss of two
shorter bands and the gain of a longer band. Each band produced by gel electrophoresis is a DNA
fragment of different size. (credit: modification of work by National Center for Biotechnology
Information)

What are the applications of DNA fingerprinting?

Solution
DNA fingerprinting:

1. DNA fingerprinting is a technique for identifying and analysing

differences in DNA between individuals.
2. It is based on DNA sequence variability and polymorphism.
3. Some of its applications include:
i. In forensic science, it is used to identify prospective
criminal suspects.
ii. To prove paternity and establish familial ties.
iii. To identify and protect the commercial crop and livestock
types.
iv. To figure out an organism's evolutionary history and the
relationships between different groupings of species.