Mention the steps involved in the separation and isolation of DNA fragments through agarose gel electrophoresis.
Or
Describe in brief the principle of DNA isolation through gel electrophoresis.
Answers
Answer:
The cutting of DNA by restriction endonucleases results in the fragments of DNA.
* These fragments are separated by a technique called gel electrophoresis.
* Since the DNA fragments are negatively charged, they can be separated by forcing them to move towards anode under an electric field through a medium/matrix.
* Most commonly used matrix is agarose, a natural polymer extracted from sea weed.
* DNA fragments separate according to their size through sieving effect provided by the agarose gel. Hence the smaller the fragment size, farther it moves.
* The separated fragments are visualized by staining them with Ethidium bromide followed by exposure to UV radiation.
* The separated bands of DNA are cut out from the agarose gel and extracted from the gel piece. This step is called elution
Gel electrophoresis is a technique used to separate DNA fragments according to their size. DNA samples are loaded into wells (indentations) at one end of a gel, and an electric current is applied to pull them through the gel. DNA fragments are negatively charged, so they move towards the positive electrode......