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OVERVIEW
• Introduction
• Historical Background
• Types of DNA profiling
• Procedure
• Applications
The most powerful and
awesome tool acquired
by humans since the
splitting of atoms”
The Time
Magazine (U.S.A)
“
INTRODUCTION
• DNA profiling (also
called DNA testing, DNA
typing, or genetic
fingerprinting) is “technology
of using tandem repeats of
individuals to identify
individuals”
Although the majority of our DNA
is the same, there are short
pieces called “microsatellites”, which repeats many
times in a person’s DNA. It’s the
pattern of repeats that are
different between individuals.

Knowing these microsatellite DNA sequences is
the basis of “DNA
WHO INVENTED
DNA
fingerprinting
university of
Leicester, UK in
1985.
In 1994,he was
knighted by Queen
Elizabeth for
services to Science
and technology.
KEY
CONCEPT
DNA profiling r finger printing
identifies people at molecular
level.
can by used for identification.
• Based on non coding regions
of DNA
•Non coding regions have
repeating DNA sequences
•Number of repeats differs
between people
•Banding pattern on a gel is a
DNA fingerprint
TYPES OF DNA
FINGERPRINTING
RFLP-based (Restriction
fragments length polymorphism)
•Original method developed by Alec
Jeffreys
DNA can be extracted from of DNA
•1st step is the Extraction
almost any human tissue.
Sources of DNA found at a
crime scene might include
blood, semen, tissue from a
deceased victim, cells in a
hair follicle, and even saliva.
DNA extracted from items of
evidence is compared to DNA
DNA EXTRACTION
METHOD
To extract DNA
, most
commonly
used detergent
is Sodium
Dodecyl Sulfate
Digestion of proteins and
lipids around the cell and it
nucleus
DNA is extracted from
sample
•DNA in the
nucleus of the
cell is
molded, folded, a
nd protected by
Proteins
•Proteinase K is
Ethanol
Precipitation
The DNA must be purified from the cell
extract.
Ethanol lowers the effective water
concentration, causing large biomolecules to interpenetrate and
aggregate. The result is a visible
precipitate at the interface, where the
ethanol is concentrated.
 DNA will rise into the alcohol layer from
Step 2.DNA cuts with the
help of Restriction
Endonuclease

DNA is cut into millions of small
fragments with the help of restriction
Endonuclease
Restriction enzymes chop DNA at
specific sequences.
the patterns occur in different places in
We are ALL a little bit different!

BIG

WAQAS’ DNA

LITTL
E
AHMED NAWAZ’
DNA
Our DNA has different sizes of pieces so
it makes a different pattern when it’s all
WAQAS
cut up ’ DNA
AHMED NAWAZ’ DNA
RFLP Analysis
Everyone has
genetic
sequences called
variable number
tandem repeats, or
VNTRs
– Everyone has
different
amounts of VNTRs
GEL
ELECTROPHORESIS
Fragments of DNA from restriction
enzyme cleavage
are separated from each other when they
migrate
through a support called an agarose gel
– It is similar to the yummy food Jell-O
gelatin
– It is actually made out of some of the
same ingredients
• The size-based separation of Molecules
GEL
ELECTROPHORESIS
The separated DNA fragments are then
drawn out of
the gel using a nylon membrane
• The nylon membrane is treated with
chemicals that
break the hydrogen bonds in DNA and
separate the
strands
• The single stranded DNA is cross linked
to the nylon
membrane
DNA having
repeat
sequences will
only be bound
to the
radioactive or
fluorescent
probe.
The final step to making a genetic
fingerprint is to place a photographic
film on top of the nylon surface. The
probes leave marks on the film
wherever they attached to the
RFLPs. Dark bands will then show up
when the film is developed, which
marks the length of the RFLPs that
were hybridized. Researchers are
then able to read the fingerprint and
match it to others. They do this by
DNA fingerprints can b used to
determine which bone fragment
belongs to which individual.
STRs & PCR
Currently, the most popular method of DNA fingerprinting
are short tandem
repeats, or STRs for short
Unlike VNTRs which analyze minisatellites that have
repeat
sequences of 9-80 base pairs, STRs use microsatellites
which have repeat sequences of
only 2-5 base pairs, introducing the “less is more”
This was a big step forward in
philosophy to the world of DNAforensic science since the
length of DNA
fingerprinting.
fragment being analyzed is short enough to be amplified
by polymerase chain reaction
(PCR), so now we are able to analyze a very small
PCR was developed in the mid 1980’s
and used the same principles that cells
use to replicate DNA to amplify the
specified region, which is usually
between 150-3,000 base pairs in length.
STEPS IN PCR OPERATION

The solution is heated to 95 C to unzip the double
helix DNA structure.
• The solution is cooled to 55 C to allow the primers
to bind to the ends of the
DNA.
• The solution is then reheated to 75 C which is the
optimal temperature for the Taq polymerase to
ADVANTAGES OF STRs OVER
RFLP/VNTR
STRs are currently the most popular type of
DNA fingerprint, since the whole
PCR process takes only a few
hours, compared to RFLP/VNTR probe
hybridization and film exposure which can
take several days. STRs can use much
smaller samples ofand quality of the DNA
DNA than
Thus, the integrity
RFLPs/VNTRs, and can factoruse partially
even with STRs than
sample is not as great a
degraded DNA to create a fingerprint
with the traditional methods of DNA
fingerprinting
DISADVANTAGE OF
STRs
The only disadvantage of the STR
approach is it is sensitive to
contaminating DNA, so usually the
STR approach is used first, followed
by a VNTR analysis if contamination
is suspected, and enough DNA is
available.
Practical Applications of
DNA
Fingerprinting
Paternity and
Maternity
Parent-child VNTR
pattern analysis
has been used to
solve standard
father-identification
cases
Criminal Identification and
Forensics
DNA isolated
from
blood, hair, skin
cells, or other
genetic evidence
left at the scene
of a crime can be
Diagnosis of Inherited
Disorders
Diagnose inherited disorders in
both prenatal and newborn
babies
These disorders may include
cystic
fibrosis, hemophilia, Huntington
's disease, familial
CONCLUSIO
N
DNA fingerprinting is the most sophisticated
way to identify living organisms.
DNA cannot easily be altered once it is left at
a
crimescene or deposited with a
mummy, which makes it a strong forensic
tool. RFLPs and VNTRs are the traditional
methods of fingerprinting DNA, which uses a
relatively large sample that uses the method
of probe hybridization to detect
polymorphisms in the

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Dna profiling

  • 1.
  • 2. OVERVIEW • Introduction • Historical Background • Types of DNA profiling • Procedure • Applications
  • 3. The most powerful and awesome tool acquired by humans since the splitting of atoms” The Time Magazine (U.S.A) “
  • 4. INTRODUCTION • DNA profiling (also called DNA testing, DNA typing, or genetic fingerprinting) is “technology of using tandem repeats of individuals to identify individuals”
  • 5. Although the majority of our DNA is the same, there are short pieces called “microsatellites”, which repeats many times in a person’s DNA. It’s the pattern of repeats that are different between individuals. Knowing these microsatellite DNA sequences is the basis of “DNA
  • 7. university of Leicester, UK in 1985. In 1994,he was knighted by Queen Elizabeth for services to Science and technology.
  • 8. KEY CONCEPT DNA profiling r finger printing identifies people at molecular level.
  • 9. can by used for identification. • Based on non coding regions of DNA •Non coding regions have repeating DNA sequences •Number of repeats differs between people •Banding pattern on a gel is a DNA fingerprint
  • 11. RFLP-based (Restriction fragments length polymorphism) •Original method developed by Alec Jeffreys DNA can be extracted from of DNA •1st step is the Extraction almost any human tissue. Sources of DNA found at a crime scene might include blood, semen, tissue from a deceased victim, cells in a hair follicle, and even saliva. DNA extracted from items of evidence is compared to DNA
  • 12. DNA EXTRACTION METHOD To extract DNA , most commonly used detergent is Sodium Dodecyl Sulfate
  • 13. Digestion of proteins and lipids around the cell and it nucleus
  • 14. DNA is extracted from sample •DNA in the nucleus of the cell is molded, folded, a nd protected by Proteins •Proteinase K is
  • 15. Ethanol Precipitation The DNA must be purified from the cell extract. Ethanol lowers the effective water concentration, causing large biomolecules to interpenetrate and aggregate. The result is a visible precipitate at the interface, where the ethanol is concentrated.  DNA will rise into the alcohol layer from
  • 16. Step 2.DNA cuts with the help of Restriction Endonuclease DNA is cut into millions of small fragments with the help of restriction Endonuclease Restriction enzymes chop DNA at specific sequences. the patterns occur in different places in
  • 17.
  • 18. We are ALL a little bit different! BIG WAQAS’ DNA LITTL E AHMED NAWAZ’ DNA
  • 19. Our DNA has different sizes of pieces so it makes a different pattern when it’s all WAQAS cut up ’ DNA AHMED NAWAZ’ DNA
  • 20. RFLP Analysis Everyone has genetic sequences called variable number tandem repeats, or VNTRs – Everyone has different amounts of VNTRs
  • 21. GEL ELECTROPHORESIS Fragments of DNA from restriction enzyme cleavage are separated from each other when they migrate through a support called an agarose gel – It is similar to the yummy food Jell-O gelatin – It is actually made out of some of the same ingredients • The size-based separation of Molecules
  • 22.
  • 23. GEL ELECTROPHORESIS The separated DNA fragments are then drawn out of the gel using a nylon membrane • The nylon membrane is treated with chemicals that break the hydrogen bonds in DNA and separate the strands • The single stranded DNA is cross linked to the nylon membrane
  • 24. DNA having repeat sequences will only be bound to the radioactive or fluorescent probe.
  • 25. The final step to making a genetic fingerprint is to place a photographic film on top of the nylon surface. The probes leave marks on the film wherever they attached to the RFLPs. Dark bands will then show up when the film is developed, which marks the length of the RFLPs that were hybridized. Researchers are then able to read the fingerprint and match it to others. They do this by
  • 26.
  • 27. DNA fingerprints can b used to determine which bone fragment belongs to which individual.
  • 28. STRs & PCR Currently, the most popular method of DNA fingerprinting are short tandem repeats, or STRs for short Unlike VNTRs which analyze minisatellites that have repeat sequences of 9-80 base pairs, STRs use microsatellites which have repeat sequences of only 2-5 base pairs, introducing the “less is more” This was a big step forward in philosophy to the world of DNAforensic science since the length of DNA fingerprinting. fragment being analyzed is short enough to be amplified by polymerase chain reaction (PCR), so now we are able to analyze a very small
  • 29. PCR was developed in the mid 1980’s and used the same principles that cells use to replicate DNA to amplify the specified region, which is usually between 150-3,000 base pairs in length. STEPS IN PCR OPERATION The solution is heated to 95 C to unzip the double helix DNA structure. • The solution is cooled to 55 C to allow the primers to bind to the ends of the DNA. • The solution is then reheated to 75 C which is the optimal temperature for the Taq polymerase to
  • 30. ADVANTAGES OF STRs OVER RFLP/VNTR STRs are currently the most popular type of DNA fingerprint, since the whole PCR process takes only a few hours, compared to RFLP/VNTR probe hybridization and film exposure which can take several days. STRs can use much smaller samples ofand quality of the DNA DNA than Thus, the integrity RFLPs/VNTRs, and can factoruse partially even with STRs than sample is not as great a degraded DNA to create a fingerprint with the traditional methods of DNA fingerprinting
  • 31. DISADVANTAGE OF STRs The only disadvantage of the STR approach is it is sensitive to contaminating DNA, so usually the STR approach is used first, followed by a VNTR analysis if contamination is suspected, and enough DNA is available.
  • 32. Practical Applications of DNA Fingerprinting Paternity and Maternity Parent-child VNTR pattern analysis has been used to solve standard father-identification cases
  • 33. Criminal Identification and Forensics DNA isolated from blood, hair, skin cells, or other genetic evidence left at the scene of a crime can be
  • 34. Diagnosis of Inherited Disorders Diagnose inherited disorders in both prenatal and newborn babies These disorders may include cystic fibrosis, hemophilia, Huntington 's disease, familial
  • 35. CONCLUSIO N DNA fingerprinting is the most sophisticated way to identify living organisms. DNA cannot easily be altered once it is left at a crimescene or deposited with a mummy, which makes it a strong forensic tool. RFLPs and VNTRs are the traditional methods of fingerprinting DNA, which uses a relatively large sample that uses the method of probe hybridization to detect polymorphisms in the