scBS-Seq
Single-Cell Bisulfite Sequencing
scBS-seq is a version of the well-established bisulfite sequencing (BS-seq) and post-bisulfite adapter tagging (PBAT) protocols, modified to detect methylated cytosines in gDNA from single cells (Smallwood et al., 2014). In this method, after single cells are isolated, gDNA is treated with sodium bisulfite, which fragments the DNA. The converted DNA then undergoes random priming several times and is PCR-amplified for sequencing. Deep sequencing provides single-nucleotide resolution mapping of methylated cytosines.
Advantages:
- Measure DNA methylation of up to 48% (10.1 M) CpG sites within a single cell
Disadvantages:
- DNA degradation after bisulfite step (Yong et al., 2016)
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
Pisanic T. R., 2nd, Athamanolap P. and Wang T. H. Defining, distinguishing and detecting the contribution of heterogeneous methylation to cancer heterogeneity. Semin Cell Dev Biol. 2016;
Yong W. S., Hsu F. M. and Chen P. Y. Profiling genome-wide DNA methylation. Epigenetics Chromatin. 2016;9:26
Greenleaf W. J. Assaying the epigenome in limited numbers of cells. Methods. 2015;72:51-56
Liang J., Cai W. and Sun Z. Single-cell sequencing technologies: current and future. J Genet Genomics. 2014;41:513-528
Plongthongkum N., Diep D. H. and Zhang K. Advances in the profiling of DNA modifications: cytosine methylation and beyond. Nat Rev Genet. 2014;15:647-661
References:
Smallwood S. A., Lee H. J., Angermueller C., Krueger F., Saadeh H., et al. Single-cell genome-wide bisulfite sequencing for assessing epigenetic heterogeneity. Nat Methods. 2014;11:817-820