Nucleotide sequencing via repetitive single molecule hybridization
First Claim
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1. A method of analyzing nucleic acids comprising:
- (a) hybridizing one or more copies of a test oligonucleotide comprising at least two informative region sequences to a single molecule of target oligonucleotides under conditions permitting hybridization of the test oligonucleotide to a perfectly complementary sequence;
(b) measuring a signal of said hybridization of the test oligonucleotide to the target oligonucleotide, thereby completing a first hybridization trial;
(c) dehybridizing the test oligonucleotide;
(d) exposing the target oligonucleotide to one or more copies of at least one additional test oligonucleotide, said additional test oligonucleotide comprising at least two informative regions;
(e) measuring a signal from the hybridization of the additional test oligonucleotide to the target oligonucleotide, thereby completing a further hybridization trial;
(f) repeating steps c) to e) until the information content from the hybridization trials exceeds the information content of the target oligonucleotide; and
(g) analyzing said signals.
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Abstract
Methods of obtaining sequence information about target oligonucleotides by repetitive single molecule hybridization are disclosed. The methods include exposing a target oligonucleotide to one or more copies of a test oligonucleotide; measuring hybridization; dehybridizing the test oligonucleotide; and repeating until the information content from the hybridization trials equals or exceeds the information content of the target oligonucleotide.
70 Citations
26 Claims
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1. A method of analyzing nucleic acids comprising:
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(a) hybridizing one or more copies of a test oligonucleotide comprising at least two informative region sequences to a single molecule of target oligonucleotides under conditions permitting hybridization of the test oligonucleotide to a perfectly complementary sequence; (b) measuring a signal of said hybridization of the test oligonucleotide to the target oligonucleotide, thereby completing a first hybridization trial; (c) dehybridizing the test oligonucleotide; (d) exposing the target oligonucleotide to one or more copies of at least one additional test oligonucleotide, said additional test oligonucleotide comprising at least two informative regions; (e) measuring a signal from the hybridization of the additional test oligonucleotide to the target oligonucleotide, thereby completing a further hybridization trial; (f) repeating steps c) to e) until the information content from the hybridization trials exceeds the information content of the target oligonucleotide; and (g) analyzing said signals. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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14. A method of analyzing nucleic acids comprising:
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(a) hybridizing one or more copies of a test oligonucleotide to a single molecule of target oligonucleotides under conditions permitting hybridization of the test oligonucleotide to a perfectly complementary sequence; (b) measuring a signal of said hybridization of the test oligonucleotide to the target oligonucleotide, thereby completing a first hybridization trial; (c) dehybridizing the test oligonucleotide; (d) exposing the target oligonucleotide to one or more copies of at least one additional test oligonucleotide; (e) measuring a signal from the hybridization of the additional test oligonucleotide to the target oligonucleotide, thereby completing a further hybridization trial; (f) repeating steps c) to e) until the information content from the hybridization trials exceeds the information content of the target oligonucleotide; and (g) analyzing said signals; and (h) analyzing an absence of said signals to eliminate oligonucleotide test members. - View Dependent Claims (15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26)
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Specification