DNA IDENTIFICATION

K GOPI

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 HOW IS A DNA PROFILE
PRODUCED?
• DNA strands chopped into fragments
• By restriction endonucleases
• Cut DNA at intron sequences (region of the
chromosome used for DNA profilling)
• Many restriction enzymes; specific for
recognition sites

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 HOW IS A DNA PROFILE
PRODUCED?
• Mini-satelite : 20-50 base sequence
repeated for 50-100 times
• Micro-satelite : 2-4 base sequence repeated
for 5-15 times
• Restriction enzymes cut the both repeated
units leaving a mixture of mini and micro
satelite

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 HOW IS A DNA PROFILE
PRODUCED?
• The fragments are separated and identified
• Gel electrophoresis

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 GEL ELECTROPHORESIS
• A variation of chromatography
• DNA fragments placed in wells in agarose gel
medium
• The medium is buffered to maintain a constant pH
• Known DNA fragments are added to aid the
identification
• The gel medium contains dye ex: ethidium
bromide which binds to the DNA fragments

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 GEL ELECTROPHORESIS
• The dye will give fluorescence effect if placed
under UV light
• DNA bands formed
• DNA samples also added with the dye- do not
bind but moves faster than DNA so that current
can be turned off before samples run out
• DNA fragments moves towards the positive anode
• Phosphate group negatively charged
• Different rate of movement (mass and charge)
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 GEL ELECTROPHORESIS
• Needs large sample of DNA
• Minimum 50 base pairs-mini satelites
• Looks like supermarket barcode
• Smaller region can be identified by using
the extension of the method

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 SOUTHERN BLOTTING
• Alkaline buffer added (denaturates DNA and
exposing the base sequences)
• Nylon filter or nitrocellulose paper is placed
over it
• Dry absorbent paper is used to draw the DNA
fragments from gel to filter
• Leaving as blots on the filter

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 REVIEW OF THE PROCESS
AGAIN
Stage 1:
• Cells are broken down
to release DNA
• If only a small amount
of DNA is available,
amplified using the
polymerase chain
reaction (PCR)

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 REVIEW OF THE PROCESS
AGAIN
Step 2:
• The DNA is cut into
fragments using
restriction enzymes.
• Each restriction enzyme
cuts DNA at a specific
base sequence.

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 REVIEW OF THE PROCESS
AGAIN
• The sections of DNA that are cut out are called
restriction fragments.

• This 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).

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 REVIEW OF THE PROCESS
AGAIN
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.

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 REVIEW OF THE PROCESS
AGAIN
• 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 and charge.

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 REVIEW OF THE PROCESS
AGAIN
• 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.

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 REVIEW OF THE PROCESS
AGAIN
Stage 4:
• The pattern of fragment distribution is then
analysed.

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 USES OF DNA PROFILING
• DNA profiling is
used to solve
crimes and medical
problems

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 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).

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BIOLOGICAL MATERIALS USED
• 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
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 SOLVING 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
• it does not prove they committed the crime
• If the profile doesn’t match the suspect then that
suspect may be eliminated from the enquiry

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 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:

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WAS THE SUSPECT AT THE
CRIME SCENE?
Suspects Blood sample Victims
Profile from crime profile
scene

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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

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 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.

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IS THIS MAN THE FATHER OF
THE CHILD?
Mother Child Man

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 INTERESTING CASE
• Helena Greenwood was a biochemist
working on DNA analysis in US
• In 1984, David Frediani broke into her
home and held her at gunpoint for several
hours and sexually assaulted her
• She persuaded him not to kill her by telling
him that she would not tell anyone

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 INTERESTING CASE
• But she gave the description to the police and
eventually the found David
• He was granted bail and set for trial in 1985
• Helena and her husband Roger moved to San
Diego
• But 3 weeks before the trial, Roger found Helena
dead in their garden strangled and beaten
• WHO WAS THE PRIME SUSPECT?

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 INTERESTING CASE
• Evidence from credit card showed that
David had been in the area just before the
murder but no forensic evidence linking
him to Helena’s body
• David stood for the original trial and was
imprisoned for 3 years

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 INTERESTING CASE
• In 1999, the San Diego police reopened a
number of unsolved cases
• To see PCR and DNA profiling help to solve
them
• Skin fragments beneath her fingernails found
DNA fingerprint matched David
• After 15 years, he was sent for life
imprisonment

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 MINI TEST
1. Explain how protein is synthesised
2. Explain how DNA profiling is done
3. List evidences that can be used to suggest evolution
4. Differentiate genetic flow and genetic drift
5. What is natural selection?
6. List types of natural selection and their examples
7. Draw the graphical illustration on types of natural selection
8. What is speciation?
9. Describe about types of reproduction isolation.

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