Circularizing nucleic acid probe able to interlock with a target sequence through catenation
First Claim
1. A method of detecting a target nucleic acid sequence in a sample, said method comprisinga) providing a detectable probe having two free nucleic acid ends which are at least partially complementary to and capable of hybridizing to two adjacent regions of said target sequence;
- b) hybridizing said probe ends to said target sequence under hybridizing conditions;
c) covalently connecting said ends of said hybridized probe with each other to form a circularized structure which interlocks with said target sequence through catenation;
d) subjecting said target sequence and said probe to non-hybridizing conditions and optionally to exonuclease activity, wherein any non-circularized probe is removed from said target sequence;
e) optionally repeating steps b) to d) one or more times; and
f) detecting the presence of said probe, wherein the presence of said probe is indicative of the presence of said target nucleic acid sequence.
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Abstract
The invention relates to a method of detecting a target nucleic acid sequence in a sample by contacting the sample with a detectable probe having ends which hybridize to two adjacent regions of the target sequence. The hybridized probe ends are then covalently connected to form a cyclized structure interlocking with the target molecule. This structure is then subjected either to non-hybridizing conditions and/or to exonuclease activity to remove any non-cyclized probes from the target sequence. The target molecule is then detected by detecting the presence of the interlocking, cantenated probe.
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Citations
46 Claims
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1. A method of detecting a target nucleic acid sequence in a sample, said method comprising
a) providing a detectable probe having two free nucleic acid ends which are at least partially complementary to and capable of hybridizing to two adjacent regions of said target sequence; -
b) hybridizing said probe ends to said target sequence under hybridizing conditions; c) covalently connecting said ends of said hybridized probe with each other to form a circularized structure which interlocks with said target sequence through catenation; d) subjecting said target sequence and said probe to non-hybridizing conditions and optionally to exonuclease activity, wherein any non-circularized probe is removed from said target sequence; e) optionally repeating steps b) to d) one or more times; and f) detecting the presence of said probe, wherein the presence of said probe is indicative of the presence of said target nucleic acid sequence. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 16, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46)
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15. A method of selectively capturing a target nucleic acid sequence to a solid support, said method comprising
a) providing a probe having two free nucleic acid ends which are at least partially complementary to and capable of hybridizing to two adjacent regions of said target sequence, and said probe being immobilized to a solid support; -
b) hybridizing said probe ends to said target sequence under hybridizing conditions; c) covalently connecting said ends of said hybridized probe with each other to form a circularized structure which interlocks with said target sequence through catentation; and d) subjecting said support and said captured target sequence to non-hybridizing conditions, wherein any non-catenated target sequence is removed from said support. - View Dependent Claims (17)
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Specification