Immobilization of DNA with nitrogen mustard–biotin conjugate for global epigenetic analysis
| Autor: | Tomomi Suda, Ryoji Kurita, Naoshi Kojima, Takaaki Kurinomaru |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Streptavidin Immobilized Nucleic Acids Biotin Enzyme-Linked Immunosorbent Assay Biochemistry Epigenesis Genetic Analytical Chemistry law.invention Mice 03 medical and health sciences chemistry.chemical_compound law Intestine Small Animals Environmental Chemistry Mechlorethamine Epigenetics Spectroscopy Polymerase chain reaction Genome Brain Sequence Analysis DNA DNA Methylation Bisulfite genomic DNA 030104 developmental biology chemistry DNA methylation 5-Methylcytosine DNA |
| Zdroj: | Analytica Chimica Acta. 1043:107-114 |
| ISSN: | 0003-2670 |
| DOI: | 10.1016/j.aca.2018.09.008 |
| Popis: | We report the quantitative analysis of 5-methylcytosine, a representative epigenetic modification in genomic DNA, with an enzyme-linked immunosorbent assay (ELISA). We synthesized a novel hetero-bifunctional linker molecule consisting of nitrogen mustard and biotin to capture DNA on the surface of biosensing devices. The molecule can successfully immobilize genomic DNA on a streptavidin coated 96-well microplate, which was then employed for immunochemical epigenetic assessment. We achieved the sensitive and quantitative detection of 5-mC in genomic DNA samples. The CpG methylation ratios obtained from our system for mouse brain and mouse small intestine genomes were 79% and 82%, respectively. These numbers are in good agreement with the previously reported methylation ratio of 75-85%, which was identified by whole genome bisulfite sequencing. Accordingly, the present technology using our novel bifunctional linker molecule provides a fast, easy, and inexpensive method for epigenetic assessment, without the need for any conventional bisulfite treatment, polymerase chain reaction (PCR), or sequencing. |
| Databáze: | OpenAIRE |
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