DNA Composition Analysis by Nuclease Digestion and HPLC

• Steven M. Wietstock ^[1]
  1. [1] Alma College

     Alma College

     City of Alma, Estados Unidos

     
• Localización: Journal of chemical education, ISSN 0021-9584, Vol. 72, Nº 10 (October), 1995, págs. 950-952
• Idioma: inglés
• Texto completo no disponible (Saber más ...)
• Resumen

  • In the field of nucleic acid research, one physical property of genomic DNA that is of use to the biochemist is the base composition of the DNA expressed in terms of the cytosine and guanosine content. Over the years, the most common method utilized for the determination of the mole fraction of G+C is the melting temperature of the DNA as determined by a spectrophotometric method. A new method for the determination of the mole fraction of G+C has been developed that requires only a sample of heat denatured DNA, a buffered nuclease P1 solution, and a reverse phase isocratic high performance liquid chromatography system. This method is discussed in detail for this article. Results are obtained in two hours time, and the need for an extensive temperature-regulated spectroscopy system is eliminated.