DNA Fingerprinting
DNA Fingerprinting
• DNA Fingerprinting is a way to identify a certain individual, rather than simply
identifying a species or a particular trait
• A technique used by scientists to distinguish between individuals of the same species
using only samples of their DNA
• The process of DNA fingerprinting was invented by Alex Jeffreys in 1985
Biological Sample used for DNA Fingerprinting
• Blood
• Hair
• Saliva
• Semen
• Body tissue cells
Steps to DNA Fingerprinting
• There are 8 steps for DNA Fingerprinting
• Step 1: Isolation of DNA
o DNA must be recovered from cells or tissue.
o Only a small amount of blood, hair, or skin is needed to isolate DNA
Steps to DNA Fingerprinting
• Step 2:Digestion of DNA by restriction endonucleases
o The DNA is cut into fragments using restriction enzymes
o Each restriction enzyme cuts DNA at a specific base sequence
o The sections of DNA that are cut out are called restriction fragments
o 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)
o Commonly used RE are (hae III, Hinf I, Alu I etc.)
o Reaction mixture is incubated overnight at 37C
Steps to DNA Fingerprinting
• Step 3: Electrophoretic separation of different fragments
• 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.
• 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.
Steps to DNA Fingerprinting
• Step 4: Transfer DNA on Nylon/Nitrocellulose membrane
o The DNA fragments are transferred to a nylon sheet by placing the sheet on the gel
and soaking them overnight by the process southern blot.
• Step5: Probing/probe labeling
o Adding radioactive or colored probes to the nylon sheet which is complementary to
target sequences
o Each probe only sticks to one or two specific places on the sheet
Steps to DNA Fingerprinting
• Step 6: Hybridization
o Labeled probe DNA should be hybridized with the complementary sequences located
on nylon membrane for the detection of position of later
o Membrane is washed to remove non specific binding and clearing of the background
• Step 7: Autoradiography
o To detect the sequences in genome bound with the hybridized radioactive probe on
membrane
o Technique involves alignment of hybridized membrane with X-ray film
o The X-ray film alter its development shows multiple no. of bands that looks like bar
codes and known as DNA fingerprints.
Steps to DNA Fingerprinting
• Step 8: Interpretation of band patterns
o Analysis of band patterns of different individuals
o Comparison of position of bands
o Computer software are also available for the analysis of DNA fingerprints.
Uses
• Diagnosis of Inherited Disorders
o Helps diagnose disorders in both prenatal and newborn babies
o Disorders may include cystic fibrosis, hemophilia, Huntington’s disease, familial
Alzheimer’s, sickle cell anemia, thalassemia, and much more.
Uses
• Crime
o Forensic science is the use of scientific knowledge in legal situations.
o The DNA profile of each individual is highly specific.
o The chances of two people having exactly the same DNA profile is 30,000 million to
1 (except for identical twins).
• Personal Identification
o The U.S. armed services are just beginning a program where they collect DNA
fingerprints from all personnel for later use, incase they need to identify casualties or
missing people
Some Other Uses of DNA fingerprinting
• Identification of carcass of tissues
• Detection of somatic mutations or cancer
• Pathogen identification
• Detection of loci controlling quantitative traits or disease resistance
• Sex determination
• Individual identification
