Gel Electrophoresis Techniques

Gel Electrophoresis

• The main purpose of gel electrophoresis is to separate and analyze mixtures of DNA, RNA, and proteins based on their size and charge.

Role of Agarose

• Agarose is used as a sieving medium in gel electrophoresis to separate molecules based on their size.
• Agarose is a polysaccharide extracted from the cell walls of red algae, and it forms a matrix of fibers that act as a molecular sieve.

Agarose Concentration and Pore Size

• The concentration of agarose affects the pore size of the gel, with higher concentrations resulting in smaller pores and lower concentrations resulting in larger pores.

Migration Speed of Molecules

• The migration speed of molecules in an agarose gel is determined by their size, with smaller molecules migrating faster than larger molecules.

Optimal Agarose Concentration

• Most agarose gels are made between 0.7% and 2% agarose because this concentration range provides an optimal pore size for separating most DNA and RNA molecules.

Agarose Toxicity

• Agarose is non-toxic and non-inflammatory, making it an ideal component for gel electrophoresis.

Gel as a Sieving Medium

• Gel is used as a sieving medium during electrophoresis because it provides a three-dimensional matrix that retards the migration of molecules based on their size, allowing for separation and analysis.