Lecture 3: Blotting and hybridisation-based Techniques

Blotting Techniques

Methods to detect specific DNA, RNA, or proteins

Southern Blotting

Identifies DNA sequences in a sample

Northern Blotting

Detects RNA sequences in a sample

Western Blotting

Identifies specific proteins in a sample

Nucleic Acid Hybridisation

Formation of DNA double helix from complementary strands

Fluorescent in situ hybridisation (FISH)

Locates genes on chromosomes using fluorescent probes

Southern Blotting - Methodology

Technique to identify specific DNA sequences in a mixture

Northern Blotting - Methodology

Process to detect RNA sequences in a sample

Western Blotting (Immunoblotting)

Identifies proteins using antibodies

Two-Dimensional (2D) gels

Electrophoresis in two directions to separate proteins

Total protein

All proteins present in a sample

Antibody detection systems

Methods to visualize antibody-antigen interactions

Microarrays

High-throughput target hybridization on a solid surface

Microarrays - Methodology

Utilizes microscopic spots of molecules for analysis