Methods for sequential DNA amplification and sequencing

US9745624B2 · kind B2 · utility

Assignee

• BRANDEIS UNIVERSITY · US

Inventors

• Lawrence J. Wangh · Waltham, US
• John Rice · Deseronto, CA
• J. Aquiles Sanchez · Framingham, US
• Kenneth Pierce · Natick, US
• Jesse Salk · Seattle, US
• Arthur Reis · Arlington, US
• Cristina Hartshorn · Dedham, US

Key dates

• Filing date: Aug 1, 2012
• Grant date: Aug 29, 2017
• Expiry date: Aug 1, 2032

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q2600/16
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Homogenous detection during or following PCR amplification, preferably LATE-PCR, utilizing fluorescent DNA dye and indirectly excitable labeled primers and probes, improves reproducibility and quantification. Low-temperature homogeneous detection during or following non-symmetric PCR amplification, preferably LATE-PCR, utilizing fluorescent DNA dye and indirectly excitable labeled mismatch-tolerant probes permits analysis of complex targets. Sequencing sample preparation methods following LATE-PCR amplifications reduce complexity and permit “single-tube” processing.