Analysis of mitochondrial DNA in microfluidic systems.
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 45 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.
Taylor
Patricia
P
Department of Electrical and Computer Engineering, 2nd Floor, ECERF Building (9107-116St.), University of Alberta, Edmonton, Alta., Canada.
Manage
Dammika P
DP
Helmle
Karmon E
KE
Zheng
Yao
Y
Glerum
D Moira
DM
Backhouse
Christopher J
CJ
D004272:DNA, Mitochondrial / Q000032:analysis*; D015246:Deoxyribonuclease EcoRI / Q000378:metabolism; D004586:Electrophoresis / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D010957:Plasmids / Q000235:genetics
D004272:DNA, Mitochondrial; D015246:Deoxyribonuclease EcoRI