Question

What is limitations of detecting Southern Blot analisys. What is alternatives for Southern blotting? What is limitation od detecting these alternative methods?

Answer 1

Southern blotting is a technique in molecular biology to detect specific DNA sequence in DNA samples.

Limitations of Southern blotting :

1. Requires large amount of targeted DNA

2. Its more expensive than most other tests

3. Time consuming

4. Complicated process and labour intensive

Alternatives for Southern blotting :

1. PCR(Polymerase Chain Reaction)

2. Gel electrophoresis

Limitations of PCR:

-It is a highly sensitive technique, any contamination of the sample can produce misleading results.

-It can only be used to detect the presence or absence of a known pathogen or gene.

-Some prior sequence information is usually necessary, to design primers to generate a PCR product.

-Specificity of the generated PCR product can be altered by nonspecific binding of primers to other similar sequences on the template DNA.

Limitations of Gel electrophoresis:

-The gel can be melted/altered when electric current is passed through it, and provide inaccurate results.