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US 6,074,823 AGranted
DNA sequencing by mass spectrometry via exonuclease degradation
AssigneeSequenom, Inc.
InventorHubert Koster
Life of this patent
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Abstract
The invention provides fast and highly accurate mass spectrometer based processes for directly sequencing a target nucleic acid (or fragments generated from the target nucleic acid), which by means of protection, specificity of enzymatic activity, or immobilization, are unilaterally degraded in a stepwise manner via exonuclease digestion and the nucleotides, derivatives or truncated sequences detected by mass spectrometry.
Claims23
A process for determining the sequence of a target nucleic acid, comprising the steps of:
- (i) obtaining multiple nucleic acid copies of the target nucleic acid, wherein the multiple copies contain at least one mass-modified nucleotide, corresponding to one of the four possible nucleotide bases;
- (ii) cleaving the multiple nucleic acid copies from a first end to a second end with an exonuclease having an activity, which is inhibited by the mass-modified nucleotide, thereby generating base-terminated nucleic acid fragments;
- (iii) identifying the nested nucleic acid fragments by mass spectrometry;
- and
- (iv) determining the sequence of the target nucleic acid from the identified nested nucleic acid fragment.
2 more elements
Dependencies are inferred from the claim text.