Two-step hybridization and capture of a polynucleotide

US6534273B2 · kind B2 · utility

Assignee

• GEN-PROBE INCORPORATED · US

Inventors

• William G. Weisburg · San Diego, US
• Jay H. Shaw · San Diego, US
• Michael M. Becker · San Diego, US
• Mehrdad Majlessi · San Diego, US
• Steven T. Brentano · Santee, US
• Kiyotada Nunomura · Nishitokyo, JP

Key dates

• Filing date: Sep 18, 2001
• Grant date: Mar 18, 2003
• Expiry date: Sep 18, 2021

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6813
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A method for capturing a target polynucleotide in a sample onto a solid support with an attached immobilized probe by using a capture probe and two different hybridization conditions that control the order of hybridization, where the first hybridization condition allows hybridization of the capture probe to the target polynucleotide, and the second hybridization condition allows hybridization of the capture probe to the immobilized probe. The method further includes amplifying the captured target polynucleotide by hybridizing at least one primer oligonucleotide to the target polynucleotide and using nucleic acid amplification that initiates from the primer oligonucleotide.