Nanopore-based single DNA molecule characterization, identification and isolation using speed bumps
AssigneeRoche Sequencing Solutions, Inc.
InventorsRandall W. Davis, Roger J. A. Chen
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Abstract
The present invention relates to a method of using nanopores to obtain sequence information of sample DNAs in ss test DNAs. The method comprises using speed bumps to stall the ss test DNAs in the nanopores at random positions of the ss test DNAs to obtain sequence information of each and every nucleotides of the sample DNAs, and to construct the whole sequences of the sample DNAs. The present invention also relates to identification and/or isolation of test DNAs having desired sequence(s) using nanopore detectors facilitated by speed bump.
Claims9
A method for sequencing a molecule comprising a plurality of nucleotides or nucleotide analogs, the method comprising:
- applying a first electric potential having a first polarity across an array of nanopore detectors, each nanopore detector comprising a pair of electrodes, a membrane disposed between the pair of electrodes, and a nanopore disposed in the membrane;
- threading a plurality of molecules through the array of nanopore detectors in a first direction based on the first polarity of the applied first electric potential;
- measuring a first electrical signal in each nanopore detector as the molecule is threaded through the nanopore in the first direction;
- applying a second electric potential having a second polarity across the array of nanopore detectors, wherein the second polarity is opposite the first polarity;
- threading the plurality of molecules through the array of nanopore detectors in a second direction based on the second polarity of the applied second electrical potential, wherein the second direction is opposite the first direction;
- applying a third electrical potential across the array of nanopore detectors, wherein the third electrical potential has a polarity that is the same as the polarity of the first electrical potential;
- threading the plurality of molecules through the array of nanopore detectors in the first direction based on the polarity of the third electrical potential;
- measuring a second electrical signal in each nanopore detector as the molecule is threaded through the nanopore in the first direction as a result of the application of the third electrical potential;
- determining a sequence for each molecule based on the first electrical signal and the second electrical signal;
- and
- applying a fourth electrical potential across the array of nanopore detectors, wherein the fourth electrical potential has a polarity that is the same as the polarity of the first electrical potential, wherein the fourth electrical potential has a magnitude that is greater than the first electrical potential that is configured to advance the molecule past a speed bump that is slowing or stalling the threading of the molecule through the nanopore.
Dependencies are inferred from the claim text.