PBAT
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)
Advantages:
- Requires only 100 ng of DNA for amplification-free WGBS of mammalian genomes
Disadvantages:
- 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
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
Yong W. S., Hsu F. M. and Chen P. Y. Profiling genome-wide DNA methylation. Epigenetics Chromatin. 2016;9:26
References:
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
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History: PBAT
Revision by sbrumpton on 2017-06-21 07:50:20 - Show/Hide
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)
Advantages:- Requires only 100 ng of DNA for amplification-free WGBS of mammalian genomes
Disadvantages:- 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
Reagents:Illumina Library prep and Array Kit SelectorReviews:Yong W. S., Hsu F. M. and Chen P. Y. Profiling genome-wide DNA methylation. Epigenetics Chromatin. 2016;9:26References:Guo F., Yan L., Guo H., et al. The Transcriptome and DNA Methylome Landscapes of Human Primordial Germ Cells. Cell. 2015;161:1437-1452Sakashita A., Kawabata Y., Jincho Y., et al. Sex Specification and Heterogeneity of Primordial Germ Cells in Mice. PLoS One. 2015;10:e0144836