Multiplex sequencing method using primers of different lengths to detect polymorphisms
US5925520A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Feb 14, 1997 |
| Grant date | Jul 20, 1999 |
| Priority date | — |
| Expiry date | Feb 14, 2017 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6869
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method is provided for determining the identity of at least two discrete single bases each adjacent to a predetermined nucleotide base sequence in a target sample having one or more types of polynucleotide chain. The method includes incorporating a capture group into the target sample and immobilizing the target sample by means of the capture group; mixing the target sample with (i) nucleotide primers which are complementary to predetermined base sequences such that they anneal to them at positions adjacent to the bases to be identified and (ii) at least two types of dideoxoy nucleoside triphosphate (ddNTPs), each type labelled with a distinguishable fluorescent group, and (iii) a nucleotide chain extending enzyme such that ddNTPs complementary to the bases to be identified are incorporated into the nucleotide primers; eluting the extending nucleotide primers, separating the types of extended nucleotide primer on basis of size or charge and identifying the ddNTPs incorporated into each type of nucleotide primer by reference to the fluorescent characteristics associated with the distinguishable groups. Primers and a test kit are provided for use in performance of the method.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.