BSAS

BSAS

BSAS is a targeted BS-Seq method that uses PCR enrichment of targeted regions and transposome-mediated library construction for rapid generation of sequencing libraries, from low (1 ng) sample input. Genomic DNA is bisulfite-converted and subjected to PCR, using primers specific for bisulfite-converted DNA. The amplicons are subjected to Nextera XT library preparation, including dual indexing. The final libraries consist of a random insert of bisulfite-converted amplified DNA, capture probes, and specific index sequences. These libraries are multiplexed and sequenced.

Pros:

• Can be applied to any genomic region from any DNA source, including tissue and cell culture.
• Rapid and highly quantitative

Cons:

• Does not cover whole genome.
• Genome and target must be known

• BSAS: Masser D. R., Berg A. S. and Freeman W. M. (2013) Focused, high accuracy 5-methylcytosine quantitation with base resolution by benchtop next-generation sequencing. Epigenetics Chromatin 6: 33

1. Sun L., Wang J., Yin X., et al. Identification of a 5-Methylcytosine Site that may Regulate C/EBPbeta Binding and Determine Tissue-Specific Expression of the BPI Gene in Piglets. Sci Rep. 2016;6:28506
2. Ou X., Thakali K. M., Shankar K., Andres A. and Badger T. M. Maternal adiposity negatively influences infant brain white matter development. Obesity (Silver Spring). 2015;23:1047-1054