DNA 5-Methylcytosine-Specific Amplification and Sequencing
DNA 5-Methylcytosine-Specific Amplification and Sequencing
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DOI：
10.1021/jacs.9b12707
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发表时间：
2020-03-11
影响因子：
15
通讯作者：
He, Chuan
中科院分区：
文献类型：
--
作者：
Liu, Chang;Cui, Xiaolong;He, Chuan
DNA 5-甲基胞嘧啶（5mC）特异性定位受到严重DNA降解和5-羟甲基胞嘧啶（5hmC）存在的阻碍，使用传统的亚硫酸盐测序方法。在这里，我们提出了一种smc特异性全基因组扩增方法（5mC- wga），通过该方法，我们在DNA扩增过程中实现了5mC的保留，从有限的输入到10 pg的范围内，5hmC信号的干扰有限，为DNA 5mC甲基组提供了高重复性和准确性。
DNA 5-methylcytosine (5mC)-specific mapping has been hampered by severe DNA degradation and the presence of 5-hydroxymethylcytosine (5hmC) using the conventional bisulfite sequencing approach. Here, we present a SmC-specific whole-genome amplification method (5mC-WGA), with which we achieved 5mC retention during DNA amplification from limited input down to 10 pg scale with limited interference from 5hmC signals, providing DNA 5mC methylome with high reproducibility and accuracy.