Chemistry

Practical course 1.1 .: Fragment isolation from the gel

Practical course 1.1 .: Fragment isolation from the gel


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Isolation of a DNA fragment from the gel

The isolation of a DNA fragment from the agarose or polyacrylamide gel is one of the techniques often used in the laboratory. Purify PCR products or isolate DNA fragments with the desired markers (e.g. antibiotic resistance) for cloning. For this purpose, the agarose gels are stained and the respective bands are cut out of the gel under UV light using a scalpel. The fragment is then obtained by melting the agarose and extracting the DNA from the mixture.

For preparative gel electrophoresis, special gel combs are used that create gel pockets for sample volumes of 50-100 µl. Depending on the method used or the manufacturer's instructions, preparative gels sometimes have to be carried out inTAE buffer, as borate (from the TBE buffer) may interfere with later purification steps.