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Southern Blot

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...

Western Blotting

Western Blotting

Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.

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Related Experiment Video

Updated: May 5, 2026

Identification of Homologous Recombination Events in Mouse Embryonic Stem Cells Using Southern Blotting and Polymerase Chain Reaction

Identification of Homologous Recombination Events in Mouse Embryonic Stem Cells Using Southern Blotting and Polymerase Chain Reaction

Published on: November 20, 2018

Genomic Southern blot analysis.

¹Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD, Australia.

Methods in Molecular Biology (Clifton, N.J.)

|November 19, 2013

Summary

This summary is machine-generated.

This protocol details genomic Southern blot analysis for detecting gene sequences in cereal genomes. It emphasizes critical steps for high sensitivity and low background, ensuring accurate results in molecular biology research.

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Last Updated: May 5, 2026

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Area of Science:

Molecular Biology
Genomics
Plant Science

Background:

Genomic Southern blot analysis is crucial for detecting specific DNA sequences.
Cereal genomes present unique challenges for DNA analysis.
Standardized protocols are needed for reliable gene detection.

Purpose of the Study:

To provide a detailed protocol for genomic Southern blot analysis in cereal genomes.
To enable the detection of transgene or endogenous gene sequences.
To ensure high sensitivity and low background for accurate results.

Main Methods:

Size fractionation of genomic DNA.
Capillary transfer to a nylon membrane.
Hybridization with a digoxigenin-labelled probe and chemiluminescent detection.

Main Results:

The protocol generates high-quality results for gene sequence detection.
Key steps include complete DNA digestion, careful membrane handling, and optimized hybridization conditions.
Successful implementation leads to high sensitivity and limited background.

Conclusions:

This protocol offers a reliable method for genomic Southern blot analysis in cereals.
Mastering critical steps ensures successful detection of gene sequences.
The method is valuable for molecular biology and genetic research in plants.