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Updated: Jun 3, 2026
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Identification of Homologous Recombination Events in Mouse Embryonic Stem Cells Using Southern Blotting and Polymerase Chain Reaction
Published on: November 20, 2018
Detection of specific DNA sequences-the southern transfer
1MRC Molecular and Cellular Cardiology Research Unit, University of Stellenbosch Medical School, Tygerberg, South Africa.
Methods in Molecular Biology (Clifton, N.J.)
|March 5, 2011
Summary
Southern blotting enables the detection and characterization of specific DNA sequences. This molecular biology technique uses DNA probes to identify target sequences after gel electrophoresis and transfer.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Accurate identification of specific DNA sequences is crucial in molecular biology.
- Existing methods may lack the specificity or sensitivity required for certain applications.
Purpose of the Study:
- To describe a technique for the detection and characterization of specific DNA sequences.
- To introduce the Southern blot method for DNA analysis.
Main Methods:
- DNA fragmentation using restriction endonucleases.
- Separation of DNA fragments by agarose gel electrophoresis.
- Denaturation of DNA, transfer to a nitrocellulose filter, and hybridization with a labeled DNA probe.
- Detection of the hybridized probe via autoradiography.
Main Results:
- The technique allows for the specific detection of DNA sequences.
- The Southern blot procedure provides a reliable method for DNA characterization.
- Successful hybridization and detection of target DNA sequences were achieved.
Conclusions:
- The Southern blot is a powerful tool for analyzing DNA.
- This method is essential for gene mapping and DNA fingerprinting.
- The technique facilitates the identification of specific DNA fragments within complex mixtures.
Related Concept Videos
02:57Southern 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...
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...
01:57Sanger Sequencing
DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
03:00Next-generation Sequencing
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.
Next-Generation Sequencing Methods
Although all next-generation methods use different technologies, they all share a set of standard features.
03:31Labeling DNA Probes
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...