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DNA Fingerprinting
Each DNA profile is Unique




                             Alec Jeffreys, 1985
Contents

Some basic facts
Introduction
History of DNA fingerprinting
Stages of DNA fingerprinting
Methods of DNA fingerprinting
Applications
  •Crime
  •Food
  •Miscellaneous
Some baSic factS
       .
  When the
  number is              On DNA sequence of
 not known,                   nucleotide
 variable, or
                           Is repeated again
irrelevant, it
is sometimes             and again but differ
     called              for person to person
  a variable
    number       When between 10 and 60 nucleotides are
    tandem       repeated – Mini- satellite
    repeats
   (VNTR)        Those with fewer are known as 
                 microsatellites or short tandem repeat
introduction
     It is
the number of
    times
  that these
   STRs are
repeated that
  produces
                 Regions of chromosomes that code for
                  Extrons contain blocks of repeated
the variations
                 proteins are called introns.
                  nucleotides        short tandem repeats
      in          (STRs)
 individuals.    Other regions that are non-coding are called
                 extrons.
introduction
     .
                    DNA fingerprinting on
                   the basis of STR doesn’t
                       give an Individual
   DNA              fingerprint but able to
fingerprint             determine the two
    ing               samples are form the
  Is also                  same person
  called      "DNA fingerprinting" is a technique that
   DNA           allows us to see differences in DNA
 profiling     fragments from one person to another
                        in more general form.
HiStory of dna
                      fingerprinting
•Up to 1984, the only method of
establishing and authenticating
personal identification was by the
fingerprint process.



•DNA fingerprinting technique was
devised in 1985 by Alec Jeffrey at
University of Leicester in England,
while working on the sequences
within myoglobin gene.
Stages of DNA Fingeprinting


    DNA is extracted from sample



•Each cell is surrounded by
cell membrane

•DNA is enclosed within the
nucleus
Stages of DNA Fingeprinting


     DNA is extracted from sample


To extract DNA , most commonly used detergent is Sodium
Dodecyl Sulfate (SDS).
                                              Digestio
                                                n of
                                              proteins
                                                and
                                               lipids
                                              around
                                              the cell
Stages of DNA Fingerprinting


        DNA is extracted from sample

•DNA in the
nucleus of the cell is
molded, folded, and
protected by Proteins

•Proteinase K is
used to break up the
proteins
Stages of DNA Fingerprinting


                Ethanol Precipitation
The DNA must be purified from the cell extract.

Ethanol lowers the effective water concentration, causing
large bio-molecules 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 the cell extract
layer.
 The protein and grease parts dissolve in the bottom, watery
layer
Stages of DNA Fingerprinting

DNA cuts with the help of Restriction
Endonuclease
DNA is cut into millions of small fragments with the help of
restriction endonucleases

Restriction enzymes chop DNA at specific sequences.

the patterns occur in different places in different individual , the
length of the fragment differs from person to person.

e.g The EcoRI restriction enzyme recognizes the following sequence

                          -C-A-A-T-T-G-
                          -G-T-T-A-A-C
Stages of DNA Fingerprinting

Separation of DNA fragments


•DNA fragments are separated
by electrophoresis

•DNA is negatively charged
so it is attracted to the positive
end of the gel.


                                  Separated DNA on the basis of gel
                                 electrophoresis
Stages of DNA Fingerprinting

  Southern blotting


•Patterns of fragments
are transferred to a nylon
membrane by a process called
Southern blotting.
Stages of DNA Fingerprinting

  DNA fingerprints compared

nylon or nitrocellulose (From
southern blot) sheet is placed under
X-ray film

The radioactive probes on the
DNA fragments expose the film

produces visible pattern of light
and dark bands which is unique to
each individual
                                       Light and dark bands of DNA when nylon
                                       film exposed to X- ray
                                        
Methods of DNA Fingerprinting


                Restriction enzyme based
                      fingerprinting

•Relatively straight forward

•When genomic DNA is cleaved with particular restriction enzymes, the
resulting fragments can contain a minisatellite (VNTR) region

•gel electrophoresis separates the digested DNA into bands based on the
length of the fragments, the pattern of bands reflects the number of
repeats in a minisatellite

•each person’s DNA will yield a unique banding pattern
Methods of DNA Fingerprinting


            PCR – based DNA fingerprinting


• By virtue of the amplification process, it is possible to begin with only
a tiny amount of DNA

• possible to fingerprint the DNA of a single cell

• Also possible to analyze old or degraded samples using PCR-based
fingerprinting

• Degraded DNA - be nearly impossible to fingerprint with restriction
analysis
Methods of DNA Fingerprinting


                Amplified fragment Length
                     Polymorphism

Come in 90’s

remains attractive because of its relatively less complicated operation
and the cost-effectiveness

use of gel in its analysis phase, there are issues of bunching of the
VTRN's, causing misidentifications in the process.
Methods of DNA Fingerprinting


             STR ( Short tandem repeats)


Mostly widely used method for comparing samples of persons

STR analyzes how many times base pairs repeat themselves on a
particular location on a strand of DNA

Has been extremely successful for use in the personal identification of
criminal suspects, paternity issues, as well as in identification of the
diseases.
Applications of DNA fingerprinting

 Mostly used to solve crime and medical, paternity problems
  •Violent murder occurred
  •Rape cases
  • Inheritance cases
  •Immigration cases

•DNA fingerprinting also found recent applications in food
industry

•Recent application in breast cancer detection
Applications of DNA Fingerprinting

                         Crime scene
• Pattern of the
DNA profile is
compared with those
of the victim and the
suspect

• profile matches the
suspect - provides
strong evidence that
the suspect was
present at crime scene
Applications of DNA fingerprinting


                  Application in
                      Food


Product identification

Genetic diversity analyses

Traceability testing
Applications of DNA fingerprinting

             Product identification.
Based on found genetic similarities or differences, a
product's identity can be established.

makes it possible to distinguish raw materials or
products from different suppliers

High-value products, such as Basmati rice and
Arabica coffee that cannot always be easily and
reliably distinguished from inferior varieties.
Applications of DNA fingerprinting

               Genetic diversity analysis
DNA fingerprints - been applied in the genetic diversity
analysis of wine cultivars

some varieties consisted of more than one genetically distinct
variety, and that varieties that had been considered distinct were
genetically identical

Such an analysis is interesting for many crops and forms the
basis for the analysis of variations in product quality.
Applications of DNA fingerprinting

                   Traceability testing



DNA fingerprinting technology can be used for traceability
testing

comparison of a fingerprint of a cow's ear and the fingerprint
of meat products will show whether the two samples originate
from the same animal
Applications of DNA fingerprinting

                  Commercial sauerkraut
                     fermentations

 Process available generates more salt as waste – disposable
problem

To avoid this, one has to study bacterial ecology – previous it
was studied by bio- chemical methods

But , DNA fingerprinting found application – shows more
complex behavior of bacterial viruses which produces waste
Applications of DNA fingerprinting

                 Commercial sauerkraut
                    fermentations

 so , on the basis of these data, one can generate new
fermentation technology – which produces less salt waste and
increases the yield
Applications of DNA fingerprinting


                Wines – use of GM yeast
                    and GM grapes

 Scientists at the Bavarian State Institute for Viticulture
and Horticulture in Würzburg, Germany, use DNA
fingerprinting to analyze wines.

Genetically modified (GM) yeast and GM vines are
undergoing field trials
Applications of DNA fingerprinting


                    False labeling issue



 Wang Jun and his group at the Chinese University of Hong
Kong used DNA fingerprinting to differentiate between Panax
ginseng (ginseng) and Panax quinquefolius (American ginseng)

price of P. quinquefolius is usually 5–10 times greater than
that of cultivated Panax ginseng
Applications of DNA fingerprinting


          Accessions ( distinct varieties) of
                      plants

 accessions are traditionally classified on the basis of
morphology, but it is suspected that numerous duplicates are
present

problem is increased by the fact that the same accessions are
often named differently in different countries
Applications of DNA fingerprinting


         Accessions ( distinct varieties) of
                     plants

 twenty-one accessions of sweet potato from the South
African genebank were compared.

RAPD and SSR techniques were used

Results show that 4 accessions which are differently named
are the genetically same – DNA fingerprinting one can use to
identify and make gene banks of world more appropriate
Applications of DNA fingerprinting


                   Lactic acid bacteria

 DNA fingerprints of lactic acid bacteria were generated by
PCR

method made it possible to identify 37 isolates from raw milk,
industrial starters and yogurt

Differentiation at species, subspecies and strain level was
possible for
 Lactobacillusdelbrueckii subsp. lactis, Lb.delbrueckii subsp bul
garicus and Str.thermophilus
Applications of DNA fingerprinting


                      Miscellaneous

• Fingerprinting has provided them to determine whether a
woman who had breast cancer at about as low as 10 cancer
cells

 In evolutionary studies, DNA fingerprinting has even
traced the origin of all of mankind to Africa

•fingerprinting can also be used to trace plants and plant
products back to their sources
concluSion
       .
One can use            DNA fingerprinting is
DNA                   very useful technique to
fingerprinting          solve many problems
                       related to crime, rape
In near era of         and murder cases when
food science to        very few evidences are
widen and                    available
know more         •DNA samples must be prepared carefully
about the
products          •Also , STR based method of DNA
                  fingerprinting is very easy and less time
                  consuming
Dna fingerprinting
Dna fingerprinting

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Dna fingerprinting

  • 1. DNA Fingerprinting Each DNA profile is Unique Alec Jeffreys, 1985
  • 2. Contents Some basic facts Introduction History of DNA fingerprinting Stages of DNA fingerprinting Methods of DNA fingerprinting Applications •Crime •Food •Miscellaneous
  • 3. Some baSic factS . When the number is On DNA sequence of not known, nucleotide variable, or Is repeated again irrelevant, it is sometimes and again but differ called for person to person a variable number When between 10 and 60 nucleotides are tandem repeated – Mini- satellite repeats (VNTR) Those with fewer are known as  microsatellites or short tandem repeat
  • 4. introduction It is the number of times that these STRs are repeated that produces Regions of chromosomes that code for Extrons contain blocks of repeated the variations proteins are called introns. nucleotides short tandem repeats in (STRs) individuals. Other regions that are non-coding are called extrons.
  • 5. introduction . DNA fingerprinting on the basis of STR doesn’t give an Individual DNA fingerprint but able to fingerprint determine the two ing samples are form the Is also same person called "DNA fingerprinting" is a technique that DNA allows us to see differences in DNA profiling fragments from one person to another in more general form.
  • 6. HiStory of dna fingerprinting •Up to 1984, the only method of establishing and authenticating personal identification was by the fingerprint process. •DNA fingerprinting technique was devised in 1985 by Alec Jeffrey at University of Leicester in England, while working on the sequences within myoglobin gene.
  • 7. Stages of DNA Fingeprinting DNA is extracted from sample •Each cell is surrounded by cell membrane •DNA is enclosed within the nucleus
  • 8. Stages of DNA Fingeprinting DNA is extracted from sample To extract DNA , most commonly used detergent is Sodium Dodecyl Sulfate (SDS). Digestio n of proteins and lipids around the cell
  • 9. Stages of DNA Fingerprinting DNA is extracted from sample •DNA in the nucleus of the cell is molded, folded, and protected by Proteins •Proteinase K is used to break up the proteins
  • 10. Stages of DNA Fingerprinting Ethanol Precipitation The DNA must be purified from the cell extract. Ethanol lowers the effective water concentration, causing large bio-molecules 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 the cell extract layer.  The protein and grease parts dissolve in the bottom, watery layer
  • 11. Stages of DNA Fingerprinting DNA cuts with the help of Restriction Endonuclease DNA is cut into millions of small fragments with the help of restriction endonucleases Restriction enzymes chop DNA at specific sequences. the patterns occur in different places in different individual , the length of the fragment differs from person to person. e.g The EcoRI restriction enzyme recognizes the following sequence -C-A-A-T-T-G- -G-T-T-A-A-C
  • 12. Stages of DNA Fingerprinting Separation of DNA fragments •DNA fragments are separated by electrophoresis •DNA is negatively charged so it is attracted to the positive end of the gel. Separated DNA on the basis of gel electrophoresis
  • 13. Stages of DNA Fingerprinting Southern blotting •Patterns of fragments are transferred to a nylon membrane by a process called Southern blotting.
  • 14. Stages of DNA Fingerprinting DNA fingerprints compared nylon or nitrocellulose (From southern blot) sheet is placed under X-ray film The radioactive probes on the DNA fragments expose the film produces visible pattern of light and dark bands which is unique to each individual Light and dark bands of DNA when nylon film exposed to X- ray  
  • 15. Methods of DNA Fingerprinting Restriction enzyme based fingerprinting •Relatively straight forward •When genomic DNA is cleaved with particular restriction enzymes, the resulting fragments can contain a minisatellite (VNTR) region •gel electrophoresis separates the digested DNA into bands based on the length of the fragments, the pattern of bands reflects the number of repeats in a minisatellite •each person’s DNA will yield a unique banding pattern
  • 16. Methods of DNA Fingerprinting PCR – based DNA fingerprinting • By virtue of the amplification process, it is possible to begin with only a tiny amount of DNA • possible to fingerprint the DNA of a single cell • Also possible to analyze old or degraded samples using PCR-based fingerprinting • Degraded DNA - be nearly impossible to fingerprint with restriction analysis
  • 17. Methods of DNA Fingerprinting Amplified fragment Length Polymorphism Come in 90’s remains attractive because of its relatively less complicated operation and the cost-effectiveness use of gel in its analysis phase, there are issues of bunching of the VTRN's, causing misidentifications in the process.
  • 18. Methods of DNA Fingerprinting STR ( Short tandem repeats) Mostly widely used method for comparing samples of persons STR analyzes how many times base pairs repeat themselves on a particular location on a strand of DNA Has been extremely successful for use in the personal identification of criminal suspects, paternity issues, as well as in identification of the diseases.
  • 19. Applications of DNA fingerprinting Mostly used to solve crime and medical, paternity problems •Violent murder occurred •Rape cases • Inheritance cases •Immigration cases •DNA fingerprinting also found recent applications in food industry •Recent application in breast cancer detection
  • 20. Applications of DNA Fingerprinting Crime scene • Pattern of the DNA profile is compared with those of the victim and the suspect • profile matches the suspect - provides strong evidence that the suspect was present at crime scene
  • 21. Applications of DNA fingerprinting Application in Food Product identification Genetic diversity analyses Traceability testing
  • 22. Applications of DNA fingerprinting Product identification. Based on found genetic similarities or differences, a product's identity can be established. makes it possible to distinguish raw materials or products from different suppliers High-value products, such as Basmati rice and Arabica coffee that cannot always be easily and reliably distinguished from inferior varieties.
  • 23. Applications of DNA fingerprinting Genetic diversity analysis DNA fingerprints - been applied in the genetic diversity analysis of wine cultivars some varieties consisted of more than one genetically distinct variety, and that varieties that had been considered distinct were genetically identical Such an analysis is interesting for many crops and forms the basis for the analysis of variations in product quality.
  • 24. Applications of DNA fingerprinting Traceability testing DNA fingerprinting technology can be used for traceability testing comparison of a fingerprint of a cow's ear and the fingerprint of meat products will show whether the two samples originate from the same animal
  • 25. Applications of DNA fingerprinting Commercial sauerkraut fermentations  Process available generates more salt as waste – disposable problem To avoid this, one has to study bacterial ecology – previous it was studied by bio- chemical methods But , DNA fingerprinting found application – shows more complex behavior of bacterial viruses which produces waste
  • 26. Applications of DNA fingerprinting Commercial sauerkraut fermentations  so , on the basis of these data, one can generate new fermentation technology – which produces less salt waste and increases the yield
  • 27. Applications of DNA fingerprinting Wines – use of GM yeast and GM grapes  Scientists at the Bavarian State Institute for Viticulture and Horticulture in Würzburg, Germany, use DNA fingerprinting to analyze wines. Genetically modified (GM) yeast and GM vines are undergoing field trials
  • 28. Applications of DNA fingerprinting False labeling issue  Wang Jun and his group at the Chinese University of Hong Kong used DNA fingerprinting to differentiate between Panax ginseng (ginseng) and Panax quinquefolius (American ginseng) price of P. quinquefolius is usually 5–10 times greater than that of cultivated Panax ginseng
  • 29. Applications of DNA fingerprinting Accessions ( distinct varieties) of plants  accessions are traditionally classified on the basis of morphology, but it is suspected that numerous duplicates are present problem is increased by the fact that the same accessions are often named differently in different countries
  • 30. Applications of DNA fingerprinting Accessions ( distinct varieties) of plants  twenty-one accessions of sweet potato from the South African genebank were compared. RAPD and SSR techniques were used Results show that 4 accessions which are differently named are the genetically same – DNA fingerprinting one can use to identify and make gene banks of world more appropriate
  • 31. Applications of DNA fingerprinting Lactic acid bacteria  DNA fingerprints of lactic acid bacteria were generated by PCR method made it possible to identify 37 isolates from raw milk, industrial starters and yogurt Differentiation at species, subspecies and strain level was possible for  Lactobacillusdelbrueckii subsp. lactis, Lb.delbrueckii subsp bul garicus and Str.thermophilus
  • 32. Applications of DNA fingerprinting Miscellaneous • Fingerprinting has provided them to determine whether a woman who had breast cancer at about as low as 10 cancer cells In evolutionary studies, DNA fingerprinting has even traced the origin of all of mankind to Africa •fingerprinting can also be used to trace plants and plant products back to their sources
  • 33. concluSion . One can use DNA fingerprinting is DNA very useful technique to fingerprinting solve many problems related to crime, rape In near era of and murder cases when food science to very few evidences are widen and available know more •DNA samples must be prepared carefully about the products •Also , STR based method of DNA fingerprinting is very easy and less time consuming