Post-bisulfite Adapter Tagging

In PBAT, bisulfite treatment precedes adaptor tagging to avoid the bisulfite-induced fragmentation of adaptor-tagged template DNAs (Miura et al., 2012). Bisulfite treatment is followed by adapter tagging and 2 rounds of random-primer extension. This procedure generates a substantial number of unamplified reads from subnanogram quantities of DNA (Smallwood et al., 2014)


  • Requires only 100 ng of DNA for amplification-free WGBS of mammalian genomes


  • Bisulfite converts unmethylated cytosines to thymidines, reducing sequence complexity, which can make it difficult to create alignments
  • SNPs where a cytosine is converted to thymidine will be missed upon bisulfite conversion
  • Bisulfite conversion does not distinguish between 5mC and 5hmC


Illumina Library prep and Array Kit Selector


Yong W. S., Hsu F. M. and Chen P. Y. Profiling genome-wide DNA methylation. Epigenetics Chromatin. 2016;9:26


Guo F., Yan L., Guo H., et al. The Transcriptome and DNA Methylome Landscapes of Human Primordial Germ Cells. Cell. 2015;161:1437-1452

Sakashita A., Kawabata Y., Jincho Y., et al. Sex Specification and Heterogeneity of Primordial Germ Cells in Mice. PLoS One. 2015;10:e0144836