Methods for generating recombined polynucleotides
US6159687A · kind A · utility
Assignee
Inventor
Key dates
| Filing date | Mar 18, 1998 |
| Grant date | Dec 12, 2000 |
| Priority date | — |
| Expiry date | Mar 18, 2018 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N15/1027
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method for in vitro construction of a library of recombined homologous polynucleotides from a number of different starting DNA templates and primers by induced template shifts during an polynucleotide synthesis is described, whereby A. extended primers are synthesized by PA1 a) denaturing the DNA templates PA1 b) annealing primers to the templates, PA1 c) extending the said primers by use of a polymerase, PA1 d) stop the synthesis, and PA1 e) separate the extended primers from the templates, B. a template shift is induced by PA1 a) isolating the extended primers from the templates and repeating steps A.b) to A.e) using the extended primers as both primers and templates, or PA1 b) repeating steps A.b) to A.e), C. this process is terminated after an appropriate number of cycles of process steps A. and B.a), A. and B.b), or combinations thereof. Optionally the polynucleotides are amplified in a standard PCR reaction with specific primers to selectively amplify homologous polynucleotides of interest.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.