Recovery of functional DNA inserts by electroendosmotic elution during gel electrophoresis.

AUTOR(ES)

Tan, H V

RESUMO

In contrast to all previous preparative electrophoresis apparatus which used a pump, electroendosmotic elution uses bound electrical charges at the end of the separating gel to generate a buffer flow. The electroendosmotic flow increased with increasing currents and decreasing buffer concentrations: its exact characteristics for the built apparatus were determined. The electroendosmotic device was able to separate two DNA fragments differing in size by only 5% with a recovery over 95%. As demonstrated in practical examples of recovery and uses of DNA inserts, up to 10 micrograms of DNA per band can be loaded at a time. The recovered DNA can be used directly for nick-translation, ligation... without further treatment. The performances of the method are expected to improve still further if the charge density and pores of the electroendosmotic medium can be "made-to-order" to provide a better flow profile of the eluting buffer.

Documentos Relacionados

• Misincorporation during DNA synthesis, analyzed by gel electrophoresis.
• Neighboring nucleotide interactions during DNA sequencing gel electrophoresis.
• Rapid DNA sequencing by horizontal ultrathin gel electrophoresis.
• Separation of very large DNA molecules by gel electrophoresis.
• Molecular sequestration stabilizes CAP-DNA complexes during polyacrylamide gel electrophoresis.