Nanopore DNA sequencing with MspA

  • Ian M. Derrington
    Department of Physics, University of Washington, Seattle, WA 98195-1560; and
  • Tom Z. Butler
    Department of Physics, University of Washington, Seattle, WA 98195-1560; and
  • Marcus D. Collins
    Department of Physics, University of Washington, Seattle, WA 98195-1560; and
  • Elizabeth Manrao
    Department of Physics, University of Washington, Seattle, WA 98195-1560; and
  • Mikhail Pavlenok
    Department of Microbiology, University of Alabama at Birmingham, Birmingham, AL 35294
  • Michael Niederweis
    Department of Microbiology, University of Alabama at Birmingham, Birmingham, AL 35294
  • Jens H. Gundlach
    Department of Physics, University of Washington, Seattle, WA 98195-1560; and

Description

<jats:p> Nanopore sequencing has the potential to become a direct, fast, and inexpensive DNA sequencing technology. The simplest form of nanopore DNA sequencing utilizes the hypothesis that individual nucleotides of single-stranded DNA passing through a nanopore will uniquely modulate an ionic current flowing through the pore, allowing the record of the current to yield the DNA sequence. We demonstrate that the ionic current through the engineered <jats:italic>Mycobacterium smegmatis</jats:italic> porin A, MspA, has the ability to distinguish all four DNA nucleotides and resolve single-nucleotides in single-stranded DNA when double-stranded DNA temporarily holds the nucleotides in the pore constriction. Passing DNA with a series of double-stranded sections through MspA provides proof of principle of a simple DNA sequencing method using a nanopore. These findings highlight the importance of MspA in the future of nanopore sequencing. </jats:p>

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