"Unveiling Genetic Mysteries: The Art of Blotting Techniques - Southern, Northern, Western - Decoding the Blueprint of Life!"

Dr. Pooja Kumari
Former Assistant Professor, Panjab University; PhD, University of Delhi

  • Molecular biology techniques to transfer biological molecules (DNA, RNA, proteins) from a gel matrix to a membrane for analysis.
  • Essential for studying molecular structure, function, and interactions.

This read will help you in understanding why we use different types of blotting techniques.

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Southern Blotting

  • Southern Blotting:

    • Purpose: Detection of specific DNA sequences.
    • Procedure: DNA fragments separated in agarose gel, transferred to a membrane, and probed with a labeled single-stranded nucleic acid.
    • Applications: Genetic fingerprinting, gene mapping, and studying gene expression.

    Northern Blotting:

    • Purpose: Analysis of RNA molecules.
    • Procedure: RNA separated in gel, transferred to a membrane, and probed with a labeled RNA or DNA probe.
    • Applications: Examining gene expression, identifying RNA species.

    Western Blotting:

    • Purpose: Detection of specific proteins.
    • Procedure: Proteins separated by gel electrophoresis, transferred to a membrane, and probed with a labeled antibody.
    • Applications: Protein identification, quantification, and characterization.

    Eastern Blotting:

    • Purpose: Detection of post-translational modifications (PTMs) on proteins.
    • Procedure: Proteins separated by gel electrophoresis, transferred to a membrane, and probed with specific antibodies for PTMs.
    • Applications: Studying protein modifications and their roles.

    Southwestern Blotting:

    • Purpose: Detection of DNA-binding proteins.
    • Procedure: DNA fragments on a membrane are probed with a labeled protein.
    • Applications: Identifying and characterizing DNA-binding proteins.

    Far-Western Blotting:

    • Purpose: Detection of protein-protein interactions.
    • Procedure: Proteins separated by gel electrophoresis, transferred to a membrane, and probed with a labeled protein as a probe.
    • Applications: Studying protein-protein interactions in complex biological systems.

    Reverse Northern Blotting:

    • Purpose: Identification of nucleic acid sequences that interact with a specific protein.
    • Procedure: Proteins on a membrane probed with labeled nucleic acid.
    • Applications: Identifying target sequences for specific proteins.

    Dot Blotting:

    • Purpose: Rapid screening of multiple samples for the presence of a specific nucleic acid or protein.
    • Procedure: Small amounts of samples spotted onto a membrane and probed with labeled nucleic acid or antibody.
    • Applications: Screening for specific molecules in a high-throughput manner.

    Advantages of Blotting Techniques:

    • Sensitivity and specificity in detecting target molecules.
    • Versatility in analyzing different bio-molecules.
    • Important tools in molecular biology research and diagnostics.

Originally published May 2, 2019

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