Single molecule sequencing with two distinct chemistry steps
US8603741B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Feb 8, 2011 |
| Grant date | Dec 10, 2013 |
| Priority date | — |
| Expiry date | Apr 1, 2032 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q2563/107
- WIPO fieldOrganic fine chemistry
- WIPO sectorChemistry
Abstract
Methods, Compositions, and Systems are provided for nucleic acid sequencing where the sequential incorporation of nucleotides uses two distinct chemical steps. A plurality of nucleotide analogs, each having a labeled leaving group at its 3′ hydroxyl can be sequentially added to a growing strand in the presence of a selective cleaving activity that cleaves the 3′ hydroxyl leaving group preferentially after it has been incorporated. The selective cleaving agent can comprise an exonuclease activity, and the exonuclease activity can be a polymerase-associated exonuclease activity. Nucleotide analogs having labels on both a cleavable polyphosphate portion and on a 3′ hydroxyl leaving group can provide signals characteristic of nucleotide analog incorporation. Systems having illumination optics, collection optics, and substrates observe signals from the labels as they are being incorporated into a growing nucleic acid strand, allowing for the sequencing of template nucleic acids.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.