Characterizing stretched polynucleotides in a synthetic nanopassage

US8748091B2 · kind B2 · utility

Assignees

• The Board of Trustees of the University of Illinois · US
• The John Hopkins University · US

Inventors

• Gregory Timp · South Bend, US
• Winston Timp · Baltimore, US
• Utkur Mirsaidov · Urbana, US
• Aleksei Aksimentiev · Urbana, US
• Jeffrey COMER · Urbana, US

Key dates

• Filing date: Dec 17, 2010
• Grant date: Jun 10, 2014
• Expiry date: Jul 29, 2032

Classification

• Technology area (CPC B): Performing Operations; Transporting
• CPC primary: B01L3/5027
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

Methods of trapping a deformed portion of a double-stranded polynucleotide in a membrane nanopassage are provided. In an aspect, the membrane has a nanopassage that defines a confine region, wherein the membrane separates a first fluid compartment from a second fluid compartment, and the nanopassage is in fluid communication with the first and second compartments. A polynucleotide is provided to the first fluid compartment and optionally a threshold voltage for the membrane and the polynucleotide is determined. A driving voltage across the membrane that is greater than the threshold voltage is applied to force a portion of the polynucleotide sequence into the nanopassage confine region, and decreased to a holding voltage bias to trap the polynucleotide portion in the nanopassage confine region. In particular, at least one nucleotide base-pair is fixably positioned in the nanopassage confine volume. In further embodiments, any of the trapping methods are used to characterize or sequence double stranded DNA.