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Basic Methods in Cellular and Molecular Biology

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Gel Purification

Overview

Gel purification is used to recover DNA fragments after electrophoretic separation. DNA recovery from an agarose gel includes three basic steps: binding, washing and eluting from a silica column. DNA is believed to bind to silica in the presence of high salt via a salt bridge. Following binding, DNA is washed of impurities and eluted under low salt conditions disrupting this interaction.

This video goes through a step-by-step, generalized procedure for cutting out a band from the gel, gel solubilization, purification through binding to a silica column, and elution of purified DNA. In addition, the presentation discusses several tips for ensuring successful gel purification, including the importance of running an agarose gel with a marker or ladder that has DNA of known sizes.

Procedure

Gel-purification is a standard procedure performed to recover desired DNA fragments from agarose gels after electrophoretic separation. After dissolving the gel fragment and running it through a specialized filter, this procedure yields DNA freed from impurities such as salts, free nucleotides and enzymes, suitable for downstream applications.

The basic principle behind DNA recovery from agarose gel involves a sequence of bind, wash, and elute steps. Once the gel is in solubilizing buffe

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