Patent ID: 9116118
Filing Date: 2015-08-25
Classification: C12Q,G01N

Abstract:
1. A method for sequencing a nucleic acid template and identifying modified bases therein comprising: providing a substrate having an upper solution above the substrate and a lower solution below the substrate, the substrate comprising a nanopore connecting the upper solution and lower solution, the nanopore sized to pass a single stranded nucleic acid; providing a voltage across the nanopore to produce a measurable current flow through the nanopore; controlling the rate of translation of a single stranded portion of the template nucleic acid through the nanopore with a processive enzyme associated with a template nucleic acid; measuring the current through the nanopore over time as it is translated through the nanopore, wherein such measuring includes measuring the current and measuring the rate of translation; determining the sequence of a portion of the template nucleic acid as it translates through the nanopore using the measured current over time; and determining the presence of modified bases in the template nucleic acid by correlating changes in the rate of translation of the nucleic acid through the nanopore to changes in the kinetics of the processive enzyme from the interaction of the modified base with the processive enzyme.