DNA sequencing by parallel oligonucleotide extensions
DCFirst Claim
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1. A method for identifying a sequence of nucleotides in a polynucleotide, the method comprising the steps of:
- (a) extending an initializing oligonucleotide along the polynucleotide by ligating an oligonucleotide probe thereto to form an extended duplex;
(b) identifying one or more nucleotides of the polynucleotide; and
(c) repeating steps (a) and (b) until the sequence of nucleotides is determined.
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Abstract
The present invention includes, in one aspect, a kit comprising one or more sets of oligonucleotide probes whose perfectly matched duplexes with complementary sequences have similar or substantially the same free energy of duplex formation.
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8 Claims
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1. A method for identifying a sequence of nucleotides in a polynucleotide, the method comprising the steps of:
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(a) extending an initializing oligonucleotide along the polynucleotide by ligating an oligonucleotide probe thereto to form an extended duplex;
(b) identifying one or more nucleotides of the polynucleotide; and
(c) repeating steps (a) and (b) until the sequence of nucleotides is determined.
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2. A kit for DNA sequence analysis, the kit comprising one or more sets of oligonucleotide probes, wherein
(i) each probe set contains at least 50 different-sequence, single-stranded oligonucleotide probes, (ii) the oligonucleotide probes of a set have a selected length of up to 12 nucleotides, and (iii) for at least one said probe set, the different-sequence, single stranded oligonucleotide probes in that set have annealing temperatures whose maximum and minimum values differ from each other by no more than 1°
Specification