PBAT
PBAT
To avoid the bisulfite-induced loss of intact sequencing templates, in post-bisulfite adapter tagging (PBAT) bisulfite treatment is followed by adapter tagging and two rounds of random primer extension. This procedure generates a substantial number of unamplified reads from as little as subnanogram quantities of DNA
Pros:
- Requires only 100 ng of DNA for amplification-free WGBS of mammalian genomes
Cons:
- 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 distingush between 5mC and 5hmC
- Farlik M., Sheffield N. C., Nuzzo A., Datlinger P., Schonegger A., et al. (2015) Single-cell DNA methylome sequencing and bioinformatic inference of epigenomic cell-state dynamics. Cell Rep 10: 1386-1397
- Guo F., Yan L., Guo H., Li L., Hu B., et al. (2015) The Transcriptome and DNA Methylome Landscapes of Human Primordial Germ Cells. Cell 161: 1437-1452
- Miura F. and Ito T. (2015) Highly sensitive targeted methylome sequencing by post-bisulfite adaptor tagging. DNA Res 22: 13-18
- Liu S., Brind'Amour J., Karimi M. M., Shirane K., Bogutz A., et al. (2014) Setdb1 is required for germline development and silencing of H3K9me3-marked endogenous retroviruses in primordial germ cells. Genes Dev 28: 2041-2055
- Okae H., Chiba H., Hiura H., Hamada H., Sato A., et al. (2014) Genome-wide analysis of DNA methylation dynamics during early human development. PLoS Genet 10: e1004868