Methods and kits for obtaining DNA end sequence information for large cloned inserts and uses thereof
US6475731B1 · kind B1 · utility
Assignee
Inventors
Key dates
| Filing date | Jul 3, 2000 |
| Grant date | Nov 5, 2002 |
| Priority date | — |
| Expiry date | Jul 3, 2020 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6869
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention provides a method of producing a cloned insert that is representative of the ends of a large segment of DNA from the genome of an organism. Specifically, the present invention provides a clone insert sequencing strategy that comprising the steps of isolating a nucleic acid molecule that is at least 20 kb, ligating the nucleic acid molecule into a plasmid vector molecule to produce a closed circle, cutting the ligated vector with a restriction endonuclease that cuts the ligated 20 kb nucleic acid molecule in at least two sites but does not cut the vector DNA, ligating the cut nucleic acid molecule to produce a deletion in the 20 kb nucleic acid molecule and determining the sequence of the ends of the nucleic acid insert and the sequence from the restriction endonuclease site.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.