Separating human DNA mixtures using denaturing high-performance liquid chromatography

Abstract

DNA mixtures represent challenging samples that are rarely amenable to direct DNA sequence analysis and many of the strategies available to separate mixtures are both labor and time intensive. Denaturing high-performance liquid chromatography is an accurate and rapid approach for the detection and scoring of mutations. It can also be used to separate DNA mixtures. The technique relies on the chromatographic separation of crosshybridization products to isolate the individual components of a mixture. By eliminating secondary amplification and excessive manipulation prior to sequencing, denaturing high-performance liquid chromatography can streamline the analysis of conditions ranging from somatic mosaicism, microchimerism and mitochondrial heteroplasmy to evidentiary material containing mixtures of DNA encountered in forensic investigations.

Keywords::

• DHPLC
• heteroduplex
• heteroplasmy
• microchimerism
• mitochondrial DNA
• mixture separation
• mutation detection
• somatic mosaicism

Acknowledgements

This material is based upon work generously supported by funding from Helen Pustmueller, the National Institute of Justice under Grant NIJ 2003-IJ-CX-K104, and by the National Science Foundation under Grants DBI-9977691 and DBI-0200484.