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US 11,427,858 B2Granted
Methods of detecting modified and unmodified DNA
AssigneesTechnion Research & Development Foundation Limited, Ramot at Tel-Aviv University Ltd.
InventorsAmit Meller, Tal Gilboa, Chen Torfstein, Yuval Ebenstein, Elmar Weinhold, Yael Michaeli-Hoch, Assaf Grunwald
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Abstract
Methods and kits for detecting the presence of at least one target DNA sequence with or without a modification in a DNA molecule are provided.
Claims20
A method of detecting the presence of at least one target modified or unmodified DNA sequence in a DNA molecule, the method comprising:
- a. contacting said DNA molecule with at least one DNA methyltransferase enzyme (MTase) in the presence of a synthetic cofactor of said MTase, wherein said enzyme differentially binds and deposits at least a detectable moiety from said synthetic cofactor on said target sequence depending on the presence of said modification,
- b. passing said contacted DNA molecule through a nanopore of membrane or film of an apparatus comprising said membrane or film comprising said nanopore, and an electrical sensor, wherein said electrical sensor is configured to detect ion flow through said nanopore, and wherein said detectable moiety is detectable as it passes through said nanopore;
- and
- c. detecting said DNA molecule as it passes through said nanopore and detecting if said detectable moiety is present as said DNA molecule passes through said nanopore, wherein said enzyme binds and deposits on modified DNA and said presence of said detectable moiety indicates the presence of said target modified DNA sequence or said enzyme binds and deposits on unmodified DNA and said presence of said detectable moiety indicates the presence of said target unmodified DNA sequence;
- thereby detecting the presence of at least one target DNA sequence with or without a modification.
2 more elements
Dependencies are inferred from the claim text.