The RRHP™ 5-hmC Library Prep Kit is the first comprehensive solution for quantitative analysis of genome-wide 5-hmC positions at single base resolution. The Reduced Representation Hydroxymethylcytosine Profiling(RRHP) system is based on blocking MspI digestion by glucosylating 5-hmC within MspI recognition sites. Shown in the schematic below, fragments lacking glucosylated-5-hmC at the adapter-ligation junction will be cleaved and not amplified by PCR. Positive selection relies on fragments containing 5-hmC being successfully amplified and analyzed.
Sample sources: Input =100 ng intact genomic DNA. DNA can be in water, TE, or low salt buffer and should be free of enzymatic inhibitors.