Tag: agarose

Tips to Reduce Run-time of Agarose Gel Electrophoresis

Usually agarose gel electrophoresis is performed at constant low voltage (5 – 10 V/cm). High voltage results in excessive heat…

Electroendosmosis in Agarose Gel Electrophoresis

In agarose gel electrophoresis, electroendosmosis (EEO) can be defined as ‘the active water transport towards cathode (negative electrode) from anode…

Recommended Agarose Gel Percentage for Efficient Separation of DNA Fragments

Agarose Gel is a porous matrix that acts as a sieve through which negatively charged linear DNA fragments migrate under an…

Applications of Agarose Gel Electrophoresis

Agarose gel electrophoresis is extensively used for the analysis of nucleic acid. Observations made by running DNA or RNA samples on agarose gel are very helpful to determine quantity and quality of nucleic acids. These powerful features make agarose gel electrophoresis an important essential step in many well known techniques such as northern and southern blotting, RFLP, RAPD, PCR, molecular cloning etc. Since agarose gel allows the elution of biomolecules, it can also be used for the purification of nucleic acids and large protein molecules and complexes. In addition, it can also be used to analyse biological processes such as apoptosis.

Agarose Gel Electrophoresis of DNA

DNA agarose gel electrophoresis is an analytical technique of analyzing and separating DNA fragments based on their size. Agarose gel…

Agarose Gel

Agarose gel can be used for the separation of nucleic acids (DNA and RNA) and high molecular weight proteins or protein complexes.

Agarose

Agarose is a useful matrix for a number of analytical and preparative techniques including gel electrophoresis, chromatography, and support matrix to immobilize enzymes and cells. Agarose is available in a variety of forms, which differ in physical properties. The two most common agarose are standard agarose and low melting agarose. Standard agarose is most commonly used for analysis of DNA and RNA. Low melting agarose is often used for preparative purposes such as elution of DNA fragments and complexes.

Protocol: Running DNA Samples in Agarose Gel

Agarose gel is placed in an electrophoresis tank filled with an electrophoresis buffer. DNA samples are mixed with DNA loading dye and loaded onto the wells of agarose gel. Electrophoresis apparatus is connected to electric supply and electrophoresis is performed at constant voltage until the desired separation among DNA fragments is achieved. After the run is over, gel is analyzed using UV-transilluminator or Gel Doc system.

Agarose Gel Preparation Process

Preparation of Agarose Gel for DNA Analysis

Agarose Gel is a gelatin-like slab, which contains small wells for loading DNA samples. It is prepared by melting agarose in a suitable electrophoresis buffer. Molten agarose is then poured into a specialized tray (casting tray), which controls the size and shape of the gel. The comb is used to create wells in agarose gel.