What is the basic principle of agarose gel electrophoresis?

Principle of Agarose Gel Electrophoresis The rate of migration is proportional to size: smaller fragments move more quickly and wind up at the bottom of the gel. DNA is visualized by including in the gel an intercalating dye, ethidium bromide. DNA fragments take up the dye as they migrate through the gel.

What are the principles of gel electrophoresis technique?

Charged molecules move through a gel when an electric current is passed across it. An electric current is applied across the gel so that one end of the gel has a positive charge and the other end has a negative charge. The movement of charged molecules is called migration. Molecules migrate towards the opposite charge.

What is the full form of TAE buffer?

Tris acetate EDTA (TAE) and tris borate EDTA (TBE) are the two most common running buffers used in nucleic acid electrophoresis. As buffers, they have a fairly constant pH and are able to conduct electricity because of their concentration of hydrogen ions.

Why TAE buffer is used in electrophoresis?

TAE buffer is added to maintain the pH of the DNA solution to neutral. Electrolysis can lead to electrolysis of water molecules and thereby release of H+ ions. These H+ ions can interact with the negatively charged DNA, neutralizing it and therefore stopping electrophoretic movement of DNA.

What are the two types of electrophoresis?

Charged macromolecules are placed in the electric field move towards the negative or positive pole based on their charge. Nucleic acid has a negative charge and therefore it migrates towards the anode. This technique is divided into two types viz slab electrophoresis and capillary electrophoresis.

How do you prepare agarose gel samples?

1. Preparation of the Gel

  1. Weigh out the appropriate mass of agarose into an Erlenmeyer flask. Agarose gels are prepared using a w/v percentage solution.
  2. Add running buffer to the agarose-containing flask. Swirl to mix.
  3. Melt the agarose/buffer mixture.
  4. Add ethidium bromide (EtBr) to a concentration of 0.5 μg/ml.

What is the purpose of the agarose gel?

Agarose gel electrophoresis is used to resolve DNA fragments on the basis of their molecular weight. Smaller fragments migrate faster than larger ones; the distance migrated on the gel varies inversely with the logarithm of the molecular weight.