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...
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| Veröffentlicht in: | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2005-08, Vol.822 (1), p.78-84 |
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| container_end_page | 84 |
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| container_title | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences |
| container_volume | 822 |
| creator | Taylor, Patricia Manage, Dammika P. Helmle, Karmon E. Zheng, Yao Glerum, D. Moira Backhouse, Christopher J. |
| description | 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. |
| doi_str_mv | 10.1016/j.jchromb.2005.05.025 |
| format | Article |
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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.</description><subject>Analysis</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Biological and medical sciences</subject><subject>Deoxyribonuclease EcoRI - metabolism</subject><subject>DNA, Mitochondrial - analysis</subject><subject>Electrophoresis</subject><subject>Electrophoresis - methods</subject><subject>Escherichia coli - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>General pharmacology</subject><subject>Genetic Vectors</subject><subject>Heteroduplex analysis</subject><subject>Humans</subject><subject>Lab-on-a-chip</subject><subject>Medical sciences</subject><subject>Microfluidic</subject><subject>Microfluidic Analytical Techniques - methods</subject><subject>Pharmacology. 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| ispartof | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences, 2005-08, Vol.822 (1), p.78-84 |
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| language | eng |
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| source | MEDLINE; ScienceDirect Journals (5 years ago - present) |
| subjects | Analysis Analytical, structural and metabolic biochemistry Biological and medical sciences Deoxyribonuclease EcoRI - metabolism DNA, Mitochondrial - analysis Electrophoresis Electrophoresis - methods Escherichia coli - metabolism Fundamental and applied biological sciences. Psychology General pharmacology Genetic Vectors Heteroduplex analysis Humans Lab-on-a-chip Medical sciences Microfluidic Microfluidic Analytical Techniques - methods Pharmacology. Drug treatments Plasmids - genetics Restriction enzyme digest |
| title | Analysis of mitochondrial DNA in microfluidic systems |
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