Transposon end compositions and methods for modifying nucleic acids
US9080211B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Oct 24, 2009 |
| Grant date | Jul 14, 2015 |
| Priority date | — |
| Expiry date | Oct 24, 2029 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q2535/122
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention provides methods, compositions and kits for using a transposase and a transposon end for generating extensive fragmentation and 5′-tagging of double-stranded target DNA in vitro, then using a DNA polymerase for generating 5′- and 3′-tagged single-stranded DNA fragments without performing a PCR amplification reaction, wherein the first tag on the 5′-ends exhibits the sequence of the transferred transposon end and optionally, an additional arbitrary sequence, and the second tag on the 3′-ends exhibits a different sequence from the sequence exhibited by the first tag. The method is useful for generating 5′- and 3′-tagged DNA fragments for use in a variety of processes, including processes for metagenomic analysis of DNA in environmental samples, copy number variation (CNV) analysis of DNA, and comparative genomic sequencing (CGS), including massively parallel DNA sequencing (so-called “next-generation sequencing.).
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.