DNA FINGERPRINTING

1. What is DNA Fingerprinting?

 DNA fingerprinting is a technique used to identify and analyse the variations in various individuals at the
level of DNA.
 It is also called as DNA typing, DNA profiling, genetic fingerprinting, genotyping or identify testing.
 It is a technique that shows the genetic makeup of living things.
 In simple words, it can be defined as a method used to identify an individual from a sample of DNA by
looking at unique patterns in their DNA.
 It has various uses, such as;
i. in forensics for criminal investigations,
ii. in paternity testing for immigration eligibility,
iii. in genealogical and medical research and
iv. For study of animal and plant populations in the field of botany, zoology and agriculture.
v. Match tissue of organ donors for transplantation
vi. Helps finding cure for the hereditary diseases
2. History:
 Almost every cell in a human body contain DNA, of which 99% are same between two human beings.
 The remaining percentage defines the differences due to the base pairs.
 There are around three million base pairs that can be different between 2 people.
 Out of these, short sequences of around 10-60 base pairs length, of repetitive DNA shows greater
variation and are called ‘Minisatellites’, which were first discovered in 1980.
 This variation is exhibited in the number of repeated or ‘stutters’ in minisatellite sequence.
 The DNA fingerprinting was first invested by Sir Alec Jeffreys in the year 1984, when he detected
variations in human DNA in form of these minisatellites.
 Each individual has a unique pattern of minisatellites except the multiple individuals from a single zygote
such as identical twins.
 Professor Jeffreys looked at the DNA fingerprint of a human family, a cow, a baboon and a tobacco plant,
and observed DNA fragments of 15-20 base pairs which are different from each one of them.
 During observing the patterns of DNA fragments from the human family, he observed that the father and
the mother had their own pattern while the child had inherited from both the father and the mother.
 In the year 1985, Professor Jeffreys and his colleagues published their first article on DNA fingerprinting
and found its various usefulness in the field of forensic science and paternity identification.
3. Principle involved in DNA fingerprinting:
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 The human genome consists of small non-coding sequences that are easily inheritable and repeatedly
present.
 This method entails locating particular DNA sequence segments known as repetitive DNA.
 These DNAs are separated from the bulk DNA and form the satellite DNA.
 The bulk DNA forms the major peaks and the satellite DNA forms the smaller peaks in density gradient
centrifugation.
 On the basis of base composition, length of the segments and number of repetitive units, the satellite
DNA can be classified into ministaellites, microsatellites, etc.
 Professor Jeffreys used a satellite DNA as a probe that showed a high degree of polymorphism, called
Variable Number of Tandem Repeats (VNTR).
 Earlier, the technique of southern blot hybridisation using radiolabelled VNTR as probe, was used.
 In the current time, the sensitivity of the technique has been increased due to the Polymerase Chain
Reaction (PCR) technique which requires only DNA from single cell to perform the analysis.
4. Steps involved in DNA fingerprinting:
 The DNA fingerprinting is carried out by utilising various enzymes and procedures.
 It includes the isolation of DNA fragment from the organic sample and by DNA sequencing techniques
the analysis is carried out to provide the results.
 The steps included in the process used by Professor Jeffreys can be seen in the picture below;
 DNA FINGERPRINTING

Fig. 1 Steps in DNA fingerprinting

 The detailed procedure is mentioned below;
 Isolation of DNA from the sample: The isolation is done from the sample to be tested. The sample can
be from any part of the body such as blood, semen, vaginal fluids, hair roots, teeth, bones, etc. The
DNA is extracted from the sample by lysis method (Phenol-chloroform DNA extraction strategy).

Fig. 2 Isolation of DNA from sample

 DNA sequencing: In many situations, there is only a small amount of sample DNA is obtained for
DNA fingerprinting. So, for these situations, DNA sequencing is carried out through Polymerase
Chain Reactions (PCRs), where multiple copies of a single DNA sample can be obtained. After this,
the DNA will undergo testing, where it will be treated with restriction endonuclease (RE) enzymes to
produce different sized fragments, called Restriction Fragment Length Polymorphism (RFLPs).
 Electrophoresis and Blotting: The different sized DNA fragments obtained are separated by the use of
an electrophoresis technique called gel electrophoresis, in which the DNA fragments are filled with
the wells of gel coated tray and electric current is applied. The DNA fragments get separated on the
basis of their size. The gel containing DNA fragments are then immersed in Sodium hydroxide
(NaOH) solution to denature DNA into single strands. These single strand DNAs are transferred to a
nylon or nitrocellulose membrane through blotting technique and are cross-linked against the
membrane by using heat or UV-light.
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Fig
. 3:

DNA sequencing and Electrophoresis

 Hybridization and Interpreting results: VNTR probes tagged with radioactive substances are now
added which form bonds with specific nucleotide sequences that are complementary to them. These
hybridised DNA shows up on photographic film as the strands of DNA decay and leave dark spots on
the film which forms the unique band called the DNA bands of a person, which eventually forms the
DNA fingerprint. This is done by autoraiography. These banding patterns are then compared with
banding patterns of the standard against which it is to be tested.

Fig. 4: Process of DNA Fingerprinting

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5. Case Studies:
 After the successful publication of the article on DNA fingerprinting, it found its various forensic utilities
and soon it has been a problem solving tool for the lawyers.
 Some of the initial case studies are mentioned below;
i. First case, Immigration Dispute:
 A family from Ghana immigrated to UK and became citizens.
 However, one of the sons went back to Ghana and was refused to returning to UK.
 In this case, Professor Jeffreys was contacted by their lawyer and an identity test was carried out.
 In this case, the samples of DNA were taken from the son, whose identity is disputed and from the
mother’s three undisputed sons along with the mother (as the father was absent).
 The patterns confirmed the relationship between the mother and the son and this shown how DNA
fingerprinting technique can be used for forensic cases and identity determination.
ii. Newer Methods and the “Angel of Death”:
 By the end of 1986, DNA fingerprinting technique was used all over the world and it was refined
by the introduction of PCR along with the discovery of microsatellites (STRs) and minisatellites
(VNTRs).
 PCR based DNA typing was used to end the hunt of 40 year for Nazi prison camp doctor, Josef
Mengele, nicknamed as “Angel of Death”, was believed to escape from the Allies of World War II
and fled to South America.
 In 1979, police found that, Mengele had drowned in the sea and was buried in Brazil.
 In 1985, the badly decomposed remains were found so that DNA samples could be taken.
 But the samples were in so poor condition that Professor Jeffreys has to carry out reverse paternity
test by using samples from Mengele’s wife and son to reconstitute the DNA pattern.
 In 1992, it was confirmed that the remains were those of Mengele.
6. Conclusion:
 DNA fingerprinting is a chemical test that reveals a person’s or any other living beings genetic makeup.
 DNA evidence is easy to obtain because genetic material is found in all human beings and thus, it has
many applications in the world.
 Twenty years after the development of DNA fingerprinting, DNA analysis remains the key to linking
suspects to biological evidence and to identify individuals in crimes and disasters.
 The list of additional use of DNA fingerprinting continues to grow and there will be discovery of new
applications using this technique.