Solid phase sequencing of biopolymers
First Claim
1. A method for detecting whether a target nucleic acid having a known sequence is present in a sample comprising nucleic acid fragments, comprising:
- (a) under hybridization conditions, contacting the nucleic acid fragments of the sample with an array of nucleic acid probes, wherein each probe in the array of nucleic acid probes comprises a double-stranded region, a single-stranded region and a variable sequence within the single-stranded region, and wherein the single-stranded region of the probes comprises a sequence complementary to a sequence of the target nucleic acid;
(b) releasing the nucleic acid fragments hybridized to the probes, thereby producing released fragments;
(c) determining molecular weights of the released fragments by mass spectrometry;
(d) determining whether the sequence of the target nucleic acid is present in the released fragments from the molecular weights of the released fragments; and
(e) detecting whether the target nucleic acid is present in the sample based on whether the target nucleic acid sequence is present in the released fragments.
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Abstract
This invention relates to methods for detecting and sequencing target nucleic acid sequences, to mass modified nucleic acid probes and arrays of probes useful in these methods, and to kits and systems which contain these probes. Useful methods involve hybridizing the nucleic acids or nucleic acids which represent complementary or homologous sequences of the target to an array of nucleic acid probes. These probes comprise a single-stranded portion, an optional double-stranded portion and a variable sequence within the single-stranded portion. The molecular weights of the hybridized nucleic acids of the set can be determined by mass spectroscopy, and the sequence of the target determined from the molecular weights of the fragments. Probes may be affixed to a solid support such as a hybridization chip to facilitate automated molecular weight analysis and identification of the target sequence.
171 Citations
15 Claims
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1. A method for detecting whether a target nucleic acid having a known sequence is present in a sample comprising nucleic acid fragments, comprising:
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(a) under hybridization conditions, contacting the nucleic acid fragments of the sample with an array of nucleic acid probes, wherein each probe in the array of nucleic acid probes comprises a double-stranded region, a single-stranded region and a variable sequence within the single-stranded region, and wherein the single-stranded region of the probes comprises a sequence complementary to a sequence of the target nucleic acid; (b) releasing the nucleic acid fragments hybridized to the probes, thereby producing released fragments; (c) determining molecular weights of the released fragments by mass spectrometry; (d) determining whether the sequence of the target nucleic acid is present in the released fragments from the molecular weights of the released fragments; and (e) detecting whether the target nucleic acid is present in the sample based on whether the target nucleic acid sequence is present in the released fragments. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15)
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Specification