Method for detecting hydroxylmethylation modification in nucleic acid and use thereof
US9567633B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Nov 21, 2012 |
| Grant date | Feb 14, 2017 |
| Priority date | — |
| Expiry date | Feb 28, 2033 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q2600/154
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A method for detecting hydroxymethylation modification in nucleic acid comprises: glycosylating the nucleic acid, digesting with MspI, ligating the digested fragments to a biotin-labeled linker at both ends thereof, digesting with NlaIII; capturing the digested fragments using streptavidin magnetic beads to produce fragments having the biotin-labeled linker at one end and a CATG 4-base sticky end at the other end, wherein these fragments reveal modification information of their adjacent CCGG sites; ligating the CATG sticky end to a linker containing a recognition site of MmeI or Ecop15I, digesting with corresponding restriction endonuclease to produce short sequence fragments that can reveal modification information of their adjacent CCGG sites; and performing a tag number comparison to obtain information about methylation and hydroxymethylation modification relative levels. A use of the method is also provided.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.