Methods for nucleic acid mapping and identification of fine-structural-variations in nucleic acids
US8329400B2 · kind B2 · utility
Assignee
Inventor
Key dates
| Filing date | Jul 10, 2009 |
| Grant date | Dec 11, 2012 |
| Priority date | — |
| Expiry date | Jun 28, 2030 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N15/66
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method of juxtaposing sequence tags (GVTs) that are unique positional markers along the length of a population of target nucleic acid molecules is provided, the method comprising: fragmenting the target nucleic acid molecule to form target DNA insert; ligating the target DNA insert to a DNA vector or backbone to create a circular molecule; digesting the target DNA insert endonuclease to cleave the target DNA insert at a distance from each end of the target DNA insert yielding two GVTs comprising terminal sequences of the target DNA insert attached to an undigested linear backbone; recircularizing the linear backbone with the attached GVTs to obtain a circular DNA containing a GVT-pair having two juxtaposed GVTs; and recovering the GVT-pair DNA by nucleic acid amplification or digestion with endonuclease having sites flanking the GVT-pair. Cosmid vectors are provided for creating GVT-pairs of ˜45- to 50-kb separation sequencable by next-generation DNA sequencers.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.