Methods for high fidelity production of long nucleic acid molecules

US8507226B2 · kind B2 · utility

Assignee

• Massachusetts Institute of Technology · US

Inventors

• Peter A. Carr · Medford, US
• Brian Yichiun Chow · Cambridge, US
• Joseph M. Jacobson · Newton, US
• David Mosley · Philadelphia, US
• Christopher J. Emig · Napa, US

Key dates

• Filing date: Jun 26, 2012
• Grant date: Aug 13, 2013
• Expiry date: Jun 26, 2032

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6848
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

In a method for synthesizing a pool of nucleic acid molecules, a first nucleic acid has a first 5′ region and a first 3′ region and a second nucleic acid has a second 5′ region and a second 3′ region. The second 3′ region and the first 5′ region have identical nucleic acid sequences. The first 3′ region is hybridized with an oligonucleotide, extending the hybridized oligonucleotide and producing a first extension product having a 3′ region complementary to the first 5′ region. The second nucleic acid is hybridized with the first extension product to hybridize the 3′ region of the first extension product to the second 3′ region, extending the 3′ region of the first extension product and producing a second extension product having a 3′ region complementary to the second 5′ region. Error-containing molecules are separated from error-free molecules by a component that selects for a sequence error.