CONTENT -Hybridization Introduction -Southern Hybridization Blotting (Transfer Of DNA On A Membrane) -Application -DNA Fingerprinting -DNA Typing Introduction-Hybridization The capacity of denatured DNA to reanneal under appropriate ionic strength and temperature is known as Hybridization. In case of a mixture of DNA molecules, hybridization leads to formation of hybrid molecules. Often a DNA probe is used in hybridization for detecting or finding a particular DNA sequence from a mixture of DNA molecules. Hybridization may be done with DNA fragments separated by electrophoresis, dot blot: DNA spots on a membrane or colony blot: hybridization with bacterial colonies containing particular rDNA. Southern Hybridization A method of DNA hybridization established by Ed Southern in 1975. Genomic DNA from an organism is restriction digested and separated by electrophoresis. The electropherosed DNA is transferred on to a membrane. The membrane bound DNA is denatured using an alkali. A DNA probe is radiolabeled and added to the denatured DNA in presence of appropriate buffer for hybridization. After sufficient time for hybridization the membrane is exposed to X-ray film. The X-ray film shows signals of hybridization: where the DNA probe has complementary structure on the transferred DNA. The resulting X-ray film is known as an Autoradiogram and the process Autoradiography. DNA fingerprinting Using the method of DNA hybridization with an appropriate DNA probe to identify individuals as done with fingerprinting. The method was first done by Sir Alec Jeffreys in 1985. Any biological material such as a drop of blood, saliva, semen, and any body part such as bones, tissue, skull, teeth, hair with root etc found at the scene of crime is used as source DNA. DNA probe used for DNA fingerprinting may be a VNTR or minisatellite DNA. The probe is labeled and used to hybridize the source DNA. An autoradiogram is generated that show different band patterns for different individuals.

1. BY: NUSRAT SHEIKH

2. CONTENT
 Hybridization Introduction
 Southern Hybridization
 Blotting (Transfer Of DNA On A Membrane)
 Application
 DNA Fingerprinting
 DNA Typing

3. Introduction-Hybridization
 The capacity of denatured DNA to reanneal under appropriate
ionic strength and temperature is known as Hybridization.
 In case of a mixture of DNA molecules, hybridization leads to
formation of hybrid molecules.
 Often a DNA probe is used in hybridization for detecting or
finding a particular DNA sequence from a mixture of DNA
molecules.
 Hybridization may be done with DNA fragments separated by
electrophoresis, dot blot: DNA spots on a membrane or colony
blot: hybridization with bacterial colonies containing particular
rDNA.

4. Southern Hybridization
 A method of DNA hybridization established by Ed Southern in 1975.
 Genomic DNA from an organism is restriction digested and
separated by electrophoresis.
 The electropherosed DNA is transferred on to a membrane.
 The membrane bound DNA is denatured using an alkali.
 A DNA probe is radiolabeled and added to the denatured DNA in
presence of appropriate buffer for hybridization.
 After sufficient time for hybridization the membrane is exposed to
X-ray film.

5.  The X-ray film shows signals of hybridization: where the DNA
probe has complementary structure on the transferred DNA.
 The resulting X-ray film is known as an Autoradiogram and the
process Autoradiography.

7. Blotting (transfer of DNA on a membrane)

8. Application Of Southern Blotting
 Identification of a single gene in a pool of DNA
fragments.
 Gene mapping.
 Analysis of genetic patterns of dna.
 Detection of specific dna sequences in a genome.
 Study of gene deletions, duplications, and mutations that
cause various diseases.

9. DNA fingerprinting
 Using the method of DNA hybridization with an appropriate DNA
probe to identify individuals as done with fingerprinting.
 The method was first done by Sir Alec Jeffreys in 1985.
 Any biological material such as a drop of blood, saliva, semen, and
any body part such as bones, tissue, skull, teeth, hair with root etc
found at the scene of crime is used as source DNA.
 DNA probe used for DNA fingerprinting may be a VNTR or
minisatellite DNA.
 The probe is labeled and used to hybridize the source DNA.
 An autoradiogram is generated that show different band patterns for
different individuals.

10. Step By Step Procedure Of Fingerprinting

11. DNA Fingerprinting Assay Sample

12. DNA Typing
 DNA typing is a procedure wherein DNA extracted from a biological
sample obtained from an individual is analyzed. The DNA is
processed to generate a pattern for each person that is generally
termed as a ' DNA profile'. This profile is unique for each person
excepting that derived from identical twins.

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