Title |
Random shearing as an alternative to digestion for mitochondrial DNA processing in droplet digital PCR
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Published in |
Mitochondrion, November 2016
|
DOI | 10.1016/j.mito.2016.11.005 |
Pubmed ID | |
Authors |
Andrej Vitomirov, Miguel Ramirez-Gaona, Sanjay R. Mehta, Josué Pérez-Santiago |
Abstract |
Droplet digital PCR (ddPCR) is a quantitative assay that requires DNA fragmentation to maximize reaction efficiency. Here, we measured the proportion of mitochondrial DNA (mtDNA) carrying the "common deletion," a rare event, to compare quantification sensitivities between alternative DNA fragmentation methods (sonication and QIAshredder spin columns) against enzymatic digestion (traditionally used). QIAshredder showed the highest sensitivity when compared to sonication, followed by digestion. Also, both sonication and QIAshredder fragmentation had shorter processing times than enzymatic digestion; therefore, QIAshredder fragmentation and sonication are alternative DNA processing methods that maximize ddPCR quantification for the detection of rare events. |
X Demographics
Geographical breakdown
Country | Count | As % |
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Unknown | 1 | 100% |
Demographic breakdown
Type | Count | As % |
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Scientists | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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China | 1 | 5% |
Unknown | 21 | 95% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 4 | 18% |
Researcher | 3 | 14% |
Unspecified | 2 | 9% |
Professor > Associate Professor | 2 | 9% |
Student > Master | 1 | 5% |
Other | 3 | 14% |
Unknown | 7 | 32% |
Readers by discipline | Count | As % |
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Agricultural and Biological Sciences | 5 | 23% |
Biochemistry, Genetics and Molecular Biology | 4 | 18% |
Unspecified | 2 | 9% |
Psychology | 1 | 5% |
Materials Science | 1 | 5% |
Other | 0 | 0% |
Unknown | 9 | 41% |