GSE61193
JBP1-seq: a fast and efficient method for genome-wide profiling of 5hmC
We developed a novel approach, J-binding protein 1 sequencing (JBP1-seq), that combines  the benefits of an improved recombinant JBP1 protein, Nextera-based library construction, and nextgeneration  sequencing (NGS) for genome-wide profiling of 5-hydroxymethylcytosine (5hmC).  Compared with the original JBP1, this new recombinant JBP1 was biotinylatedin vivo and conjugated  to magnetic beads via biotin-streptavidin interactions. These modifications allowed a more efficient  and consistent pull-down of ß-glucosyl-5-hydroxymethylcytosine (ß-glu-5hmC), and sequence-ready  libraries can be generated within 4.5 hours from DNA inputs as low as 50 ng. 5hmC enrichment of  human brain DNA using the new JBP1 resulted in over 25,000 peaks called, which is significantly  higher than the 4,003 peaks enriched using the old JBP1. Comparison of the technical duplicates and  validations with other platforms indicated the results are reproducible and reliable. Thus, JBP1-seq  provides a fast, efficient, cost-effective method for accurate 5hmC genome-wide profiling. Overall design: An improvement of JBP1-Seq