Method for producing a polynucleotide for use in single primer amplification

US5612199A · kind A · utility

Assignee

• Behringwerke Aktiengesellschaft · DE

Inventors

• Linda M. Western · Mountain View, US
• Karen M. Hahnenberger · Cupertino, US
• Samuel J. Rose · Mountain View, US
• Martin Becker · West Hollywood, US
• Edwin F. Ullman · Atherton, US

Key dates

• Filing date: Apr 1, 1994
• Grant date: Mar 18, 1997
• Expiry date: Apr 1, 2014

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6853
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A method is disclosed for extending an extender probe to produce a single stranded polydeoxynucleotide that is free of unreacted extender probe and has two segments that are non-contiguous and complementary with each other. The method comprises the steps of (1) providing in combination (a) a polynucleotide having two non-contiguous, non-complementary nucleotide sequences S1 and S2 wherein S2 is 5' of S1 and is at least ten deoxynucleotides long, (b) an extender probe comprised of two deoxynucleotide sequences, wherein the sequence at the 3'-end of the extender probe (EP1) is hybridizable with S1 and the other of the deoxynucleotide sequences (EP2) is substantially identical to S2 and (c) means for modifying the 3'-end of extender probe that does not hybridize with the polynucleotide and (2) extending the extender probe along the polynucleotide wherein extender probe not hybridized to the polynucleotide becomes modified at its 3'-end.