What is Northern and Southern blotting?
Table of Contents
What is Northern and Southern blotting?
Northern and Southern blotting are standard molecular biology techniques for identification and quantification of RNA and DNA respectively. Effective isolation and detection of RNA and DNA in molecular biology research is critical to gene discovery, sequencing, and mapping used in diagnostics and industry applications.
What is Northern blotting steps?
Steps Involved in Northern Analysis RNA isolation (total or poly(A) RNA) Probe generation. Denaturing agarose gel electrophoresis. Transfer to solid support and immobilization. Prehybridization and hybridization with probe.
Why is it called a northern blot?
Northern blotting takes its name from its similarity to the first blotting technique, the Southern blot, named for biologist Edwin Southern. The major difference is that RNA, rather than DNA, is analyzed in the northern blot.
What is the 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.
Why it is called Northern blotting?
What are the applications of northern blot?
Applications of Northern Blot Northern blotting is used as a sensitive test for the detection of transcription of DNA fragments that are to be used as a probe in Southern Blotting. It also allows the detection and quantification of specific mRNAs from different tissues and different living organisms.
Which gel is used in Northern blotting?
For northern blotting, either total or poly(A)+ RNA can be used. Most cellular RNAs are single-stranded polynucleotides and generally have secondary structures formed by intramolecular base-pairing. The secondary structures prevent RNA from being separated according to size by electrophoresis through an agarose gel.
What is Southern blot used for?
Southern blot analysis can be used to investigate whether a gene is amplified, deleted, or structurally rearranged in cancer cells as compared to normal cells. Although this technique is quite labor-intensive, it’s particularly useful for detecting large deletions in tumor genomes.
What is a northern blot used for?
Northern blot is a laboratory analysis method used to study RNA. Specifically, purified RNA fragments from a biological sample (such as blood or tissue) are separated by using an electric current to move them through a sieve-like gel or matrix, which allows smaller fragments to move faster than larger fragments.
