What are the applications of Southern blotting?

Applications of Southern Blotting

• Identifying specific DNA in a DNA sample.
• Preparation of RFLP (Restriction Fragment Length Polymorphism) maps.
• Detection of mutations, deletions or gene rearrangements in DNA.
• For criminal identification and DNA fingerprinting (VNTR)

What are the application of blotting techniques?

Blotting methods are the adjunct to gel electrophoresis, a method for separating DNA, RNA and proteins with exceptional resolving power. It can be applied to biomolecules that will adhere stably to a support material such as nitrocellulose, nylon or paper membrane and are still able to bind their cognate ligand.

What is the key difference between Southern and Northern blotting?

While both techniques are used to identify nucleic acid sequences, Northern blotting is performed to detect RNA sequences, while Southern blotting is done to detect DNA sequences. The processes for each are similar, involving gel electrophoresis, transfer to a membrane, and hybridization.

Why it is called northern blotting?

The term ‘northern blot’ actually refers specifically to the capillary transfer of RNA from the electrophoresis gel to the blotting membrane. Northern blotting takes its name from its similarity to the first blotting technique, the Southern blot, named for biologist Edwin Southern.

What are the advantages and disadvantages of northern blotting?

northern blots: pros & cons. The procedure is relatively fast, “low- tech” and cheap. Standard Northern procedure is, in general, less sensitive than nuclease protection assays and RT-PCR. Approximately 100,000 copies of a DNA or RNA sequence are required for detection by blot hybridization.

What is principle of Southern blotting?

Principle. Southern blotting is based on the principle of separation of DNA fragments by gel electrophoresis followed by the identification by labeled probe hybridization. The DNA fragments are separated based on their size and charge during electrophoresis.

What kind of probe is used in Southern blotting?

The membrane is then treated with a small piece of DNA or RNA called a probe, which has been designed to have a sequence that is complementary to a particular DNA sequence in the sample; this allows the probe to hybridize, or bind, to a specific DNA fragment on the membrane.

What are the advantages of northern blotting?

The advantages of using northern blotting include the detection of RNA size, the observation of alternate splice products, the use of probes with partial homology, the quality and quantity of RNA can be measured on the gel prior to blotting, and the membranes can be stored and reprobed for years after blotting.

When to use a northern or Southern blot?

Northern blotting is primarily used to: Identify the specific RNA sequence in a given sample. It helps in disease diagnosis. Image 5: The image shows the procedures involved in a southern blotting method.

Why are Southern blots used in DNA testing?

Southern blotting method is used for the following reasons: To detect a specific DNA sequence in a given sample. It is used in paternity testing. It is used in DNA fingerprinting. It is used for victim identification. It is used for identifying criminals. It is useful in genetic diseases and infectious agents.

How are Northwestern and Southwestern blotting techniques used?

To study genetic changes in DNA; it can be used for homology-based cloning too. Northwestern blotting studies that detect interactions between RNA and proteins. Southwestern blotting studies that detect DNA-binding proteins. The two techniques stem from the same principle and differ in the type of molecules they target.

Who is the inventor of the Southern blotting technique?

It was developed by Edward M. Southern (1975). Southern blotting is a hybridization technique for identification of particular size of DNA from the mixture of other similar molecules.