Methods, kits and compositions pertaining to PNA Molecular Beacons
First Claim
1. A method for in-situ analysis of a target sequence of an organism of interest in a sample, said method comprising:
- a) providing a sample of fixed cells for in-situ analysis;
b) treating the sample with a polymer of covalently linked subunits, said polymer comprising;
i) a non-polynucleotide probing nucleobase sequence that is complementary or substantially complementary to a target sequence;
ii) a first arm segment and a second arm segment that are covalently linked to, or embedded within, the probing nucleobase sequence and wherein at least one of the first or second arm segments is covalently linked to the probing nucleobase sequence through a flexible linkage;
iii) at least one linked donor moiety and at least one linked acceptor moiety, wherein said donor and acceptor moieties are covalently linked to the polymer at positions that are separated by at least a portion of the probing nucleobase sequence;
c) detecting, identifying or quantitating the hybridization of the polymer to the target sequence, under suitable in-situ hybridization conditions, wherein the presence, absence or amount of target sequence present in the sample is correlated with a change in detectable signal associated with at least one donor or acceptor moiety of the polymer; and
d) detecting, identifying or quantitating the organism of interest in the sample based upon the presence, absence or amount of the hybridization of the polymer to the target sequence that is determined.
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Accused Products
Abstract
This invention is directed to methods, kits and compositions pertaining to PNA Molecular Beacons. PNA Molecular Beacons comprise self-complementary arm segments and flexible linkages that promote intramolecular or intermolecular interactions. In the absence of a target sequence, PNA Molecular Beacons facilitate efficient energy transfer between the linked donor and acceptor moieties of the probe. Upon hybridization of the probe to a target sequence, there is a measurable change in at least one property of at least one donor or acceptor moiety of the probe which can be used to detect, identify or quantitate the target sequence in a sample.
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Citations
7 Claims
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1. A method for in-situ analysis of a target sequence of an organism of interest in a sample, said method comprising:
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a) providing a sample of fixed cells for in-situ analysis;
b) treating the sample with a polymer of covalently linked subunits, said polymer comprising;
i) a non-polynucleotide probing nucleobase sequence that is complementary or substantially complementary to a target sequence;
ii) a first arm segment and a second arm segment that are covalently linked to, or embedded within, the probing nucleobase sequence and wherein at least one of the first or second arm segments is covalently linked to the probing nucleobase sequence through a flexible linkage;
iii) at least one linked donor moiety and at least one linked acceptor moiety, wherein said donor and acceptor moieties are covalently linked to the polymer at positions that are separated by at least a portion of the probing nucleobase sequence;
c) detecting, identifying or quantitating the hybridization of the polymer to the target sequence, under suitable in-situ hybridization conditions, wherein the presence, absence or amount of target sequence present in the sample is correlated with a change in detectable signal associated with at least one donor or acceptor moiety of the polymer; and
d) detecting, identifying or quantitating the organism of interest in the sample based upon the presence, absence or amount of the hybridization of the polymer to the target sequence that is determined. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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Specification