Circularized templates for sequencing
US10752944B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Sep 6, 2012 |
| Grant date | Aug 25, 2020 |
| Priority date | — |
| Expiry date | Oct 5, 2036 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6855
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The invention provides methods of forming a circular template for sequencing a target nucleic acid. The circular template is generated by amplification of a segment of the target nucleic acid with chimeric primers with complementary 5′ ends. The circular template has a single nick or gap providing a site for initiation of template-directed extension for sequence analysis. Sequencing of a single template generates reads of alternating segments of the same strand of the target nucleic spaced by primer segments. The different reads of the same strand of the target nucleic acid can be compiled to generate a consensus sequence. Because only one strand of the target nucleic acid is sequenced per reaction, the present method avoids errors introduced by unwittingly combining sequences of both strands of a heteroduplex PCR product. Because only one strand of the target nucleic acid is sequenced per reaction, the present method avoids errors introduced by unwittingly combining sequences of both strands of a heteroduplex PCR product.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.