DNA sequencing by mass spectrometry via exonuclease degradation
First Claim
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1. A method of determining a sequence of a nucleic acid, comprising the steps of:
- (i) obtaining multiple copies of the nucleic acid to be sequenced;
(ii) cleaving the multiple copies from a first end to a second end with an exonuclease to sequentially release individual nucleotides;
(iii) identifying each of the sequentially released nucleotides by mass spectrometry; and
(iv) determining the sequence of the nucleic acid from the identified nucleotides.
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Abstract
Methods for determining the sequence of nucleic acids by cleaving the nueleic acid unilaterally from a first end with an exonuclease activity to sequentially release individual nucleotides, identifying each of the sequentially release nucleotides by mass spectrometry, and determining the sequence of the nueleic acid from the identified nucleotides are disclosed. The method is amenable to multiplexing for simultaneously determining more than one nucleic acid sequence.
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Citations
28 Claims
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1. A method of determining a sequence of a nucleic acid, comprising the steps of:
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(i) obtaining multiple copies of the nucleic acid to be sequenced; (ii) cleaving the multiple copies from a first end to a second end with an exonuclease to sequentially release individual nucleotides; (iii) identifying each of the sequentially released nucleotides by mass spectrometry; and (iv) determining the sequence of the nucleic acid from the identified nucleotides. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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20. A method of determining a sequence of a nucleic acid, comprising the steps of:
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(i) obtaining multiple copies of the nucleic acid to be sequenced; (ii) cleaving the multiple copies from a first end to a second end with an exonuclease to produce multiple sets of nested nucleic acid fragments; (iii) determining the molecular weight value of each one of the sets of nucleic acid fragments by mass spectrometry; and (iv) determining the sequence of the nucleic acid from the molecular weight values of the sets of nucleic acid fragments. - View Dependent Claims (21, 22, 23, 24, 25, 26, 27, 28)
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Specification