Systematic evolution of ligands by exponential enrichment: Solution SELEX
US5567588A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Jun 5, 1995 |
| Grant date | Oct 22, 1996 |
| Priority date | — |
| Expiry date | Jun 5, 2015 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N2333/976
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
Described herein are methods for improved partitioning between high and low affinity nucleic acid ligands identified through the SELEX method, termed solution SELEX. The solution SELEX method achieves partitioning between high and low affinity nucleic acid-target complexes through a number of methods, including (1) primer extension inhibition which results in differentiable cDNA products. Primer extension inhibition is achieved with the use of nucleic acid polymerases, including DNA or RNA polymerases, reverse transcriptase, and Q.beta.-replicase; (2) exonuclease hydrolysis inhibition which results in only the highest affinity ligands amplifying during PCR. This is achieved with the use of any 3'.fwdarw.5' double-stranded exonuclease; (3) linear to circle formation to generate molecules amplifiable during PCR; or (4) PCR amplification of single-stranded nucleic acids. A central theme of the method of the present invention is that the nucleic acid candidate mixture is screened in solution and results in preferential amplification of the highest affinity RNA ligand or catalytic RNA.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.