| Autor: |
Larkin J; Department of Physics, Northeastern University, Boston, Massachusetts 02115, USA., Henley RY; Department of Physics, Northeastern University, Boston, Massachusetts 02115, USA., Jadhav V; Department of Physics, Northeastern University, Boston, Massachusetts 02115, USA., Korlach J; Pacific Biosciences, Menlo Park, California 94025, USA., Wanunu M; Department of Physics, Northeastern University, Boston, Massachusetts 02115, USA.; Department of Chemistry and Chemical Biology, Northeastern University, Boston, Massachusetts 02115, USA. |
| Abstrakt: |
Compared with conventional methods, single-molecule real-time (SMRT) DNA sequencing exhibits longer read lengths than conventional methods, less GC bias, and the ability to read DNA base modifications. However, reading DNA sequence from sub-nanogram quantities is impractical owing to inefficient delivery of DNA molecules into the confines of zero-mode waveguides-zeptolitre optical cavities in which DNA sequencing proceeds. Here, we show that the efficiency of voltage-induced DNA loading into waveguides equipped with nanopores at their floors is five orders of magnitude greater than existing methods. In addition, we find that DNA loading is nearly length-independent, unlike diffusive loading, which is biased towards shorter fragments. We demonstrate here loading and proof-of-principle four-colour sequence readout of a polymerase-bound 20,000-base-pair-long DNA template within seconds from a sub-nanogram input quantity, a step towards low-input DNA sequencing and mammalian epigenomic mapping of native DNA samples. |
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