2024年9月20日 · 2021年，发表在 Genome Research 上的一项研究提出一个基于酶反应的EM-seq技术 [1]，该技术可以检测DNA上的5mC和5hmC修饰，且不引入通常与亚硫酸氢盐测序相关的偏差。 该技术主要需要3种酶和2次反应。 01.TET2： 甲基胞嘧啶双加氧酶2（Tet methylcytosine dioxygenase 2） 02.T4-BGT： T4噬菌体-糖基转移酶（T4-phage beta-glucosyltransferase) 03.APOBEC3A： 载脂蛋白B mRNA编辑酶催化亚基3A（apolipoprotein B mRNA editing …

EM-seq libraries displayed even GC distribution, better correlations across DNA inputs, increased numbers of CpGs within genomic features, and accuracy of cytosine methylation calls. EM-seq was effective using as little as 100 pg of DNA, and these libraries maintained the described advantages over bisulfite sequencing.

NEBNext Enzymatic Methyl-seq (EM-seq™) is a new method for identification of 5-mC and 5-hmC. While bisulfite sequencing has been the gold standard for methylome analysis, this conversion treatment damages DNA, resulting in fragmentation, loss and bias.

NEBNext Enzymatic Methyl-seq (EM-seq ™) is a high-performance enzyme-based alternative to bisulfite conversion for the identification of 5mC and 5hmC. Unlike bisulfite conversion, this highly efficient method minimizes DNA damage, resulting in superior detection of methylated cytosines, with fewer sequencing reads.

EM-seq可实现纳克级别DNA微量建库，实现单碱基分辨率的DNA甲基化鉴定，可作为癌症早期筛查与诊断、疾病预后监测、个性化医疗评估、器官移植后监测等临床研究的有力工具。

近年来，基于酶法转化的甲基化测序技术不断涌现出来，APOBEC偶联甲基化表观测序 (ACE-seq）可以特异性检测5hmC，以及酶法甲基化测序(EM-seq)可同时检测5mC和5hmC。

近年来，基于酶法转化的甲基化测序技术不断涌现出来，APOBEC偶联甲基化表观测序 (ACE-seq）可以特异性检测5hmC，以及酶法甲基化测序(EM-seq)可同时检测5mC和5hmC。

将EM-seq与RNA-seq、ATAC-seq、NOMe-seq、NicE-seq、Hi-C、组蛋白修饰和ChIP-seq等数据结合起来进行多组学比较，可以更完整的理解细胞调控。 与WGBS相比，EM-seq可实现以单碱基分辨率检测基因组甲基化，并且不破坏DNA，更加温和。 另外在基因组覆盖度、CpG检测率和基因组特征识别等方面都得到了改进。 EM-seq可以用于较低的起始量以及降解DNA的检测。

The NEBNext Enzymatic Methyl-seq Kit (EM-seq™) combines NEBNext® Ultra™ II reagents with these two enzymatic steps to construct Illumina® libraries that accurately represent 5mC and 5hmC within the genome. Converted libraries are amplified using …

EM-seq (Enzymatic Methyl-sequencing) is a powerful technique that has been developed to study DNA methylation patterns with high precision and sensitivity. EM-sep, based on enzymatic treatment of DNA, utilizes specific enzymes that can selectively modify or …