Increasing the accuracy of nanopore DNA sequencing using a time-varying cross membrane voltage
Autor: | Henry Brinkerhoff, Kenji Doering, Matthew T. Noakes, Jonathan W. Mount, Jasmine Bowman, Jens H. Gundlach, Andrew H. Laszlo, Katherine S. Baker, Benjamin I. Tickman, Kyle W. Langford, Ian M. Derrington |
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Rok vydání: | 2019 |
Předmět: |
Materials science
Sequence analysis Mycobacterium smegmatis Biomedical Engineering Porins Bioengineering Applied Microbiology and Biotechnology Signal Nanopores 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine A-DNA 030304 developmental biology 0303 health sciences biology DNA Helicases Helicase DNA Sequence Analysis DNA Nanopore chemistry biology.protein Molecular Medicine Base calling Nanopore sequencing Biological system Algorithms 030217 neurology & neurosurgery Biotechnology |
Zdroj: | Nature Biotechnology. 37:651-656 |
ISSN: | 1546-1696 1087-0156 |
Popis: | Nanopore DNA sequencing is limited by low base-calling accuracy. Improved base-calling accuracy has so far relied on specialized base-calling algorithms, different nanopores and motor enzymes, or biochemical methods to re-read DNA molecules. Two primary error modes hamper sequencing accuracy: enzyme mis-steps and sequences with indistinguishable signals. We vary the driving voltage from 100 to 200 mV, with a frequency of 200 Hz, across a Mycobacterium smegmatis porin A (MspA) nanopore, thus changing how the DNA strand moves through the nanopore. A DNA helicase moves the DNA through the nanopore in discrete steps, and the variable voltage moves the DNA continuously between these steps. The electronic signal produced with variable voltage is used to overcome the primary error modes in base calling. We found that single-passage de novo base-calling accuracy of 62.7 ± 0.5% with a constant driving voltage improves to 79.3 ± 0.3% with a variable driving voltage. The variable-voltage sequencing mode is complementary to other methods to boost the accuracy of nanopore sequencing and could be incorporated into any enzyme-actuated nanopore sequencing device. The accuracy of nanopore DNA sequencing is substantially improved by application of a variable-voltage sequencing mode. |
Databáze: | OpenAIRE |
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