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Rapid and affordable genome-wide bisulfite DNA sequencing by XmaI-reduced representation bisulfite sequencing.
- Source :
-
Epigenomics [Epigenomics] 2017 Jun; Vol. 9 (6), pp. 833-847. Date of Electronic Publication: 2017 May 10. - Publication Year :
- 2017
-
Abstract
- Aim: To develop a reduced representation bisulfite sequencing (RRBS) approach for rapid and affordable genome-wide DNA methylation analysis.<br />Methods: We have selected restriction endonuclease XmaI to produce RRBS library fragments. After digestion and partial fill-in DNA fragments were ligated to barcoded adapters, bisulfite converted, size-selected, and sequenced on the Ion Torrent Personal Genome Machine. XmaI-RRBS results were compared with the previously published RRBS data.<br />Results: We have developed an XmaI-RRBS method for rapid and affordable genome-wide DNA methylation analysis, with library preparation taking only 4 days and sequencing possible within 4 h. We have also addressed several challenges in order to further improve the RRBS technology. XmaI-RRBS may be performed on degraded DNA samples and is compatible with the bench-top next-generation sequencing machines.
- Subjects :
- Deoxyribonucleases, Type II Site-Specific chemistry
High-Throughput Nucleotide Sequencing economics
High-Throughput Nucleotide Sequencing standards
Humans
MCF-7 Cells
Whole Genome Sequencing economics
Whole Genome Sequencing standards
DNA Methylation
High-Throughput Nucleotide Sequencing methods
Whole Genome Sequencing methods
Subjects
Details
- Language :
- English
- ISSN :
- 1750-192X
- Volume :
- 9
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Epigenomics
- Publication Type :
- Academic Journal
- Accession number :
- 28488887
- Full Text :
- https://doi.org/10.2217/epi-2017-0031