UNIT 10.4 Agarose Gel Electrophoresis

Abstract

Agarose gel electrophoresis is a simple and highly effective method for separating, identifying, and purifying 0.5- to 25-kb DNA fragments. The Basic Protocol in this unit can be divided into three stages: (1) a gel is prepared with an agarose concentration appropriate for the size of DNA fragments to be separated; (2) the DNA samples are loaded into the sample wells and the gel is run at a voltage and for a time period that will achieve optimal separation; and (3) the gel is stained or, if ethidium bromide has been incorporated into the gel and electrophoresis buffer, visualized directly upon illumination with UV light. Support protocols describe the use of midigels and minigels, as well as recommendations for photographing stained gels.