Reagents for reversibly terminating primer extension

US8212020B2 · kind B2 · utility

Inventors

• Steven Albert Benner · Gainesville, US
• Daniel Hutter · Gainesville, US
• Nicole A Leal · Gainesville, US
• Fei Chen · Thạnh Trung, VN

Key dates

• Filing date: Mar 23, 2009
• Grant date: Jul 3, 2012
• Expiry date: Mar 13, 2030

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C07H23/00
• WIPO field: Organic fine chemistry
• WIPO sector: Chemistry

Abstract

This invention relates to the field of nucleic acid chemistry, more specifically to the field of compositions of matter that comprise triphosphates of modified 2′-deoxynucleosides and oligonucleotides that are formed when these are appended to the 3′-end of a primer, wherein said modifications comprise NH2 moiety attached to their 3′-hydroxyl group and a fluorescent species in a form of a tag affixed to the nucleobase via a linker that can be cleaved. Such compositions and their associated processes enable and improve the sequencing of oligonucleotides using a strategy of cyclic reversible termination, as outlined in U.S. Pat. No. 6,664,079. Most specifically, the invention concerns compositions of matter that are 5′-triphosphates of ribo- and 2′-deoxyribonucleosides carrying detectable tags and oligonucleotides that might be derived from them. The invention also concerns processes wherein a DNA polymerase, RNA polymerase, or reverse transcriptase synthesizes said oligonucleotides via addition of said triphosphates to a primer.