Southern blotting technology
Blot hybridization is the process of binding a nucleic acid fragment to be tested to a certain solid phase support and then hybridizing it to a labeled probe present in the liquid phase.Southern blot is the process of transferring DNA to a solid phase support and then hybridizing it to a labeled probe present in the liquid phase.
Southern blotting detects the content of DNA molecules of a specific size, and is commonly used for enzymatic profiling of cloned genes, qualitative and quantitative analysis of genomic genes, mutation analysis, and restriction length polymorphism (RELP) analysis.
Principle
The basic principle of Southern blotting technology is to first separate DNA fragments digested by restriction endonuclease by gel electrophoresis, then denature the DNA on the gel and transfer the single-stranded DNA fragments in situ onto a nitrocellulose membrane or other solid-phase support.
By labeling a specific DNA probe with a radioactive isotope, performing a hybridization reaction with the single-stranded DNA fragment, and analyzing the hybridization signals by radioactive autoradiography to determine the position of each DNA band complementary to the probe, the position and size of the DNA fragment containing a particular sequence among the numerous enzymatic digestion products can be determined.
Appliance
Southern blotting technology can detect the content of DNA molecules of specific sizes, and is commonly used for enzymatic profiling of cloned genes, qualitative and quantitative analysis of genomic genes, mutation analysis, and restriction length polymorphism (RELP) analysis.
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