Polymerase chain reaction amplification method using a single primer which randomly anneals
US5665572A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Aug 23, 1994 |
| Grant date | Sep 9, 1997 |
| Priority date | — |
| Expiry date | Aug 23, 2014 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/686
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method of amplifying template DNA by polymerase chain reaction (PCR) in which a single oligonucleotide primer having a restriction site is contacted with the template DNA, whereby the oligonucleotide randomly anneals to a single strand of the template DNA and DNA sequences complementary to the single strand are synthesized. An initial PCR amplification yields synthetic DNA sequences having the oligonucleotide sequence incorporated therein at the 5' end, and a sequence complementary to the template DNA. A second PCR amplification under higher stringency conditions amplifies regions of the template DNA to give DNA fragments having the restriction sites of the oligonucleotide primer. Thereby the method can be used to amplify trace quantities of template DNA of unknown sequence simply and efficiently, which has applications in the construction of DNA libraries of chromosome specific regions and the development of probes for chromosome mapping.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.