Method for automated molecular cloning
US6420144B1 · kind B1 · utility
Assignee
Inventors
Key dates
| Filing date | Jun 28, 2000 |
| Grant date | Jul 16, 2002 |
| Priority date | — |
| Expiry date | Jun 28, 2020 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/686
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method for the automated molecular cloning and amplification of closed circular clones in vitro in a cell-free system using a pair of particularly designed cloning primers in a ligation-during-amplification (LDA) process, in which the fully extended DNA strands containing the donor sequence are ligated by DNA ligase and then used as templates for subsequent amplification. By designing particular cloning primers, the present invention can be used to obtain amplified closed circular nucleic acid clones with inverted or non-inverted insertions or substitutions with a donor nucleic acid fragment, or deletions of a fragment of the closed circular nucleic acid.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.