PB_seq
Protein/DNA Binding Followed by High-Throughput Sequencing
PB_seq is a DNA-binding assay that allows the DNA-protein binding energy landscape to be characterized genome-wide, in the absence of chromatin (Guertin et al., 2012). It belongs to the family of methods more commonly known as systematic evolution of ligands by exponential enrichment (SELEX) (Ozer et al., 2014). Genomic DNA is sonicated, size-selected, and purified. After hybridization to the DNA-binding protein the protein-bound DNA is isolated, extracted, and prepared for sequencing.
Advantages:
- Determines binding efficiencies independent of chromatin structure
Disadvantages:
- Not yet adopted widely by the scientific community
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
None available yet
References:
Guertin M. J., Martins A. L., Siepel A. and Lis J. T. Accurate prediction of inducible transcription factor binding intensities in vivo. PLoS Genet. 2012;8:e1002610