J-Binding Protein 1 Sequencing

JBP1-seq is a method for genome-wide profiling of 5hmC. It relies on the strong affinity of the J-binding protein 1 (JBP1) for glucosylated 5hmC. The method uses a recombinant JBP1 protein with an additional His tag and Avi tag that is biotinylated and subsequently conjugated to streptavidin-coated magnetic beads (Cui et al., 2014). The JBP1-seq workflow involves 4 main steps. 1) The gDNA is tagmented to fragment it and add adapter sequences. 2) The 5hmC sites are glucosylated by T4-beta-GT. 3) The JBP1-magnetic beads are used to enrich for DNA fragments containing beta-glu-5hmC. 4) The enriched library is amplified to add P5 and P7 adapters and barcodes for sequencing.


  • Streamlined protocol


  • Regions that contain high levels of 5hmC can be overrepresented
  • Optimal resolution is approximately 50 bp


Illumina Library prep and Array Kit Selector


Shull A. Y., Noonepalle S. K., Lee E. J., Choi J. H. and Shi H. Sequencing the cancer methylome. Methods Mol Biol. 2015;1238:627-651

Plongthongkum N., Diep D. H. and Zhang K. Advances in the profiling of DNA modifications: cytosine methylation and beyond. Nat Rev Genet. 2014;15:647-661


Cui L., Chung T. H., Tan D., Sun X. and Jia X. Y. JBP1-seq: a fast and efficient method for genome-wide profiling of 5hmC. Genomics. 2014;104:368-375