Methods of amplifying nucleic acid sequences mediated by transposase/transposon DNA complexes

US10894980B2 · kind B2 · utility

Assignee

• President and Fellows of Harvard College · US

Inventors

• Chongyi Chen · Somerville, US
• Dong Xing · Laishui, CN
• Xiaoliang Sunney Xie · Lexington, US

Key dates

• Filing date: Jul 15, 2016
• Grant date: Jan 19, 2021
• Expiry date: Sep 15, 2036

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6865
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Methods are provided for nucleic acid amplification including contacting a double stranded nucleic acid with transposases bound to transposon DNA, wherein the transposon DNA includes a transposase binding site and an RNA polymerase promoter sequence, wherein the transposases/transposon DNA complex bind to target locations along the double stranded nucleic acid and cleave the double stranded nucleic acid into a plurality of double stranded fragments, with each double stranded fragment having the transposon DNA bound to each 5′ end of the double stranded fragment, extending the double stranded fragments along the transposon DNA to make double stranded extension products having double stranded RNA polymerase promoter sequences at each end, contacting the double stranded extension products with an RNA polymerase to make a plurality of RNA transcripts of each double stranded extension product, reverse transcribing the RNA transcripts into single stranded copy DNA, forming complementary strands to the single stranded copy DNA to form a plurality of double stranded DNA amplicons corresponding to each double stranded fragment.