Preparation of 13C/15N-labeled oligomers using the polymerase chain reaction
US6258567A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | May 29, 1999 |
| Grant date | Jul 10, 2001 |
| Priority date | — |
| Expiry date | May 29, 2019 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6844
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Preparation of .sup.13 C/.sup.15 N-labeled DNA oligomers using the polymerase chain reaction (PCR). A PCR based method for uniform (.sup.13 C/.sup.15 N)-labeling of DNA duplexes is described. Multiple copies of a blunt-ended duplex are cloned into a plasmid, each copy containing the sequence of interest and restriction Hinc II sequences at both the 5' and 3' ends. PCR using bi-directional primers and uniformly .sup.13 C/.sup.15 N-labeled dNTP precursors generates labeled DNA duplexes containing multiple copies of the sequence of interest. Twenty-four cycles of PCR, followed by restriction and purification, gave the uniformly .sup.13 C/.sup.15 N-labeled duplex sequence with a 30% yield. Such labeled duplexes find significant applications in multinuclear magnetic resonance spectroscopy.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.