Overview of Southern blotting, steps and application.

1. Southern Blotting
Yogesh Bhandari

2. Introduction
Discovered in 1975 by Edwin
Mellor Southern.
Used in molecular biology for
DNA analysis
Also known as Southern Blotting
and DNA Blotting

3. What is Southern Blotting
Southern blotting is a process that transfer the DNA fragments that
separated by gel electrophoresis into nitrocellulose membrane.

4. Principle
This is method of Hybridization:
It is process of forming double stranded DNA molecule between a
single stranded DNA probe and a single stranded trarget DNA.

5. Types of Blotting
• Southern Blotting
• Northern Blotting
• Western Blotting
• Eastern Blotting

6. Steps in Southern Blotting
• Collection of sample -
Extract the DNA from the cell
• Restirction Digestion -
Digest the DNA with Restriction Endonuclease

7. • Gel electrophoresis -
Mixture of fragment subjected to gel elecrophoresis
Gel separate the fragments according to size
• Denaturation -
Restriction fragment present in the gel are denatured with alkaline

8. • Blotting -
Transfer the DNA on the nitrocellulose membrane.
• Hybridization and Visualization -
Hybridization of single strand of DNA with probe
V isualization by autoradiography

10. • To identify specific DNA in a DNA sample.
• This can be applied in studying structure of gene or elucidate
restriction enzyme maps.
• It can be used in the identification of the methylated sites in case of
some genes.
Applications

11. Thank You