Determination of base (nucleotide) composition in DNA oligomers by mass spectrometry
US6613509B1 · kind B1 · utility
Assignee
Inventor
Key dates
| Filing date | Mar 22, 1999 |
| Grant date | Sep 2, 2003 |
| Priority date | — |
| Expiry date | Mar 27, 2020 |
Classification
- Technology area (CPC Y)Emerging Cross-Sectional Technologies
- CPC primaryY10S435/81
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The determination of base (nucleotide) composition in DNA by mass spectrometry is described. Accurate and efficient analyses of the enormous pool of DNA sequences are required for; (a) validation of DNA sequences; (b) comparison of a parent (known) sequence with a related (unknown) sequence, and (c) characterization of sequence polymorphisms in various genes especially those associated with genetically inherited human diseases. The combination of stable isotope-labeling of PCR products of target sequences with analysis of the mass shifts by mass spectrometry (MS) is shown to provide such analyses, since the mass-shift due to the labeling of a single type of nucleotide (i.e., A, T, G, or C) identifies the number of that type of nucleotide in a given DNA fragment. Accurate determinations of nucleotide compositions of DNA fragments have been achieved with an accuracy of ±0.03% with respect to their known sequences. The method has also been applied to identify a known single-nucleotide polymorphism (SNP). The comparisons of nucleotide compositions determined according to the teachings of the present method among homologous sequences are useful in sequence validation, sequence co…
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