Jump-seq: Capture and amplify 5hmC Sites Whole Genome-Wide. Jump-seq: Capture and amplify 5hmC Sites Whole Genome-Wide

We developed an efficient and precise method, which we name as Jump-seq, to capture and amplify 5hmC signal whole genome-wide, bridge the gap between base resolution (TAB-seq) and economic cost (hmC-seal). Overall design: The principle is to label 5hmC by 6-N3-Glucose moiety and a hairpin DNA oligonucleotide carrying an alkyne group can be covalently connected to the azide-modified 5hmC via Huisgen cycloaddition (click) chemistry. The primer extension from the hairpin motif extends to the modified 5hmC site and will continue to “land” on the genomic DNA. 5hmC sites could be inferred from the DNA polymerase “landing” pattern at the 5-prime juncture.本研究开发了一种高效精准的全基因组捕获扩增5-羟甲基胞嘧啶（5hmC）信号的方法，将其命名为Jump-seq，该方法填补了碱基分辨率级检测技术（TAB-seq）与低成本检测方案（hmC-seal）之间的技术鸿沟。实验设计原理：其核心机制为通过6-叠氮基葡萄糖（6-N3-Glucose）修饰标记5hmC，再借助携带炔基的发夹型脱氧核糖核苷酸寡聚体，通过Huisgen环加成（点击化学）与叠氮基修饰的5hmC发生共价连接。以该发夹结构为引物进行延伸反应，延伸至修饰后的5hmC位点后，可继续在基因组DNA上“着陆”；研究人员可通过DNA聚合酶在5'接合端的“着陆”模式，推断出5hmC的位点信息。