Methods of Detecting Target Nucleic Acids
AssigneeApplied Biosystems, LLC
InventorsMuhammad A. SHARAF, Timothy WOUDENBERG, Khairuzzaman Bashar MULLAH
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Abstract
The present disclosure relates to methods of identifying target nucleic acids by using coded molecules and its analysis by translocation through a nanopore. Generally, coded molecules are subject to a target polynucleotide dependent modification. The modified coded molecule is detected by isolating the modified coded molecules from the unmodified coded molecules prior to analysis through the nanopore or by detecting a change in the signal pattern of the coded molecule when analyzed through the nanopore.
Claims21
1-73. (canceled)
A method of detecting a plurality of target polynucleotides, the method comprising:
- a) contacting a population of coded molecules with a plurality of target polynucleotides, wherein the population of coded molecules comprises a plurality of subpopulations and each coded molecule of each subpopulation comprises:
- (i) one or more block polymer regions and a target probe capable of hybridizing to a target polynucleotide, wherein the target probe of each subpopulation hybridizes to a different target polynucleotide;
- and
- (ii) a detectable signal pattern distinguishable amongst the plurality of subpopulations;
- b) modifying coded molecules of the population of coded molecules by modifying the target probe with a modifying agent following hybridizing the target probe with the different target polynucleotide, wherein the modified target probe is indicative of the different target polynucleotide;
- c) translocating the modified coded molecules through a nanopore and detecting the signal pattern;
- and
- d) associating the detected signal pattern to a specific subpopulation of coded molecules in the plurality of subpopulations, wherein a difference in the detected signal pattern compared to the signal pattern of the specific subpopulation indicates the presence of the different target polynucleotide.
Dependencies are inferred from the claim text.