Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
9753281 | Journal of Chromatography B | 2005 | 7 Pages |
Abstract
Abnormalities in mitochondrial function play a major role in many human diseases. It is often of critical importance to ascertain what proportion of the mitochondria within a cell, or cells, bear a given mutation (the mitochondrial “demographics”). In this work, a rapid, novel, on-chip procedure was used, in which a restriction enzyme was employed to excise a mitochondrial DNA (mtDNA) sequence from plasmid DNA that acted as a prototypical mitochondrial genome. The DNA was then denatured, reassembled to form duplexes, fluorescently labelled and analysed. This method was able to differentiate between a homogeneous population and a heterogeneous population. Using a microfluidic chip, the method could be performed in about 45Â min, even without robotics or multiplexed operation, whereas conventional methods of analysis require days to perform. This method may ultimately form the basis for a means of characterizing the mitochondrial demographics of a single cell.
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Patricia Taylor, Dammika P. Manage, Karmon E. Helmle, Yao Zheng, D. Moira Glerum, Christopher J. Backhouse,