We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Reading DNA at single-nucleotide resolution with a mutant MspA nanopore and phi29 DNA polymerase.
- Authors
Manrao, Elizabeth A; Derrington, Ian M; Laszlo, Andrew H; Langford, Kyle W; Hopper, Matthew K; Gillgren, Nathaniel; Pavlenok, Mikhail; Niederweis, Michael; Gundlach, Jens H
- Abstract
Nanopore technologies are being developed for fast and direct sequencing of single DNA molecules through detection of ionic current modulations as DNA passes through a pore's constriction. Here we demonstrate the ability to resolve changes in current that correspond to a known DNA sequence by combining the high sensitivity of a mutated form of the protein pore Mycobacterium smegmatis porin A (MspA) with phi29 DNA polymerase (DNAP), which controls the rate of DNA translocation through the pore. As phi29 DNAP synthesizes DNA and functions like a motor to pull a single-stranded template through MspA, we observe well-resolved and reproducible ionic current levels with median durations of ∼28 ms and ionic current differences of up to 40 pA. Using six different DNA sequences with readable regions 42-53 nucleotides long, we record current traces that map to the known DNA sequences. With single-nucleotide resolution and DNA translocation control, this system integrates solutions to two long-standing hurdles to nanopore sequencing.
- Publication
Nature biotechnology, 2012, Vol 30, Issue 4, p349
- ISSN
1546-1696
- Publication type
Journal Article
- DOI
10.1038/nbt.2171