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首页> 外文会议>ASMS Conference on Mass Spectrometry and Allied Topics >A Nanoflow LC-MS/MS Approach to Monitor Rapid Loss of 5mC and 5hmC from Differentiating Erythoid Progenitor Genomes
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A Nanoflow LC-MS/MS Approach to Monitor Rapid Loss of 5mC and 5hmC from Differentiating Erythoid Progenitor Genomes

机译：纳米射线LC-MS / MS方法，以监测5MC和5HMC的快速损失，从差异分化红斑祖细胞基因组

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摘要

A highly sensitive nanoLC-MS assay was developed for 5hmC, 5mC, and dC analysis, measuring attomole quantities of digested DNA from erythroid progenitor cell lines. Analytes were detected as low as 120 amol on-column for 5hmC. The loss of pentose sugar moieties during insource fragmentation was optimized to monitor the aglycones for an enhanced sensitivity. The results generated by the assay were in agreement with those by dot-blot analysis. Both show that the percent of 5hmC (0.2%) in the genome of early progenitors rapidly declines to less than 0.05% as progenitors initiate erythoid transcription. The results suggest that 5hmC contributes to the loss of 5mC from erythoid enhancers and from LINE1 elements but that other mechanisms also contribute to global loss of 5mC.
机译：开发了高灵敏的纳米-SMS测定为5HMC，5MC和DC分析，测量来自红细胞祖细胞系的消化DNA的抗摩托数量。将分析物检测到50AMOL柱5HMC。优化insource碎片期间戊糖部分的损失被优化，以监测糖蜜增强的敏感性。测定产生的结果与点印迹分析一致。随着祖细胞剂引发红外转录，均显示出早期祖细胞基因组的5HMC（0.2％）的百分比百分比（0.2％）迅速下降至小于0.05％。结果表明，5HMC从红细胞增强剂和Line1元素中有助于5MC的损失，但其他机制也有助于全球损失5MC。

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