纳米孔在DNA中的五种C5-cytosine变体之间进行区分
Zachary L Wescoe1, Jacob Schreiber, Mark Akeson
1Department of Biomolecular Engineering, Baskin School of Engineering, MS SOE2, University of California , Santa Cruz, California 95064, United States.
Journal of the American Chemical Society
|October 28, 2014
概括
这项研究通过准确识别五种细胞因子变异来推进DNA测序,包括像5-甲基细胞因子 (mC) 和5-基细胞因子 (caC) 这样的修饰基,这对于理解表观遗传学至关重要.
科学领域:
- 生物化学 生物化学
- 基因组学就是基因组学.
- 表观遗传学 在表观遗传学中,表观遗传学是指表观遗传学.
背景情况:
- 使用酶进行纳米孔测序使单核酸DNA读取成为可能.
- 建立了区分正规,5-甲基 (mC) 和5-基甲基 (hmC).
- 哺乳动物细胞在转化5-基甲基细胞素 (hmC) 过程中产生5-基甲基细胞素 (caC) 和5-甲基细胞素 (fC).
研究的目的:
- 开发一种方法来准确识别DNA中的细胞因子及其关键C5变体 (mC,hMC,caC,fC).
- 评估使用纳米孔测序在CG二核酸中区分这些基的准确性.
主要方法:
- 利用phi29 DNA聚合酶通过M2MspA纳米孔进行受控的DNA转位.
- 使用补丁放大器记录离子电流痕迹.
- 基于三个连续的离子电流状态实现的决策边界用于基调.
主要成果:
- 在CG二核酸中实现了准确的基调用细胞因子,mC,hMC,caC和fC.
- 单通读取精度从91.6%到98.3%不等.
- 准确度受到围绕CG二核酸的最近邻基的影响.
结论:
- 纳米孔测序与酶控制相结合,可以可靠地区分多个细胞因子变异.
- 这种方法通过包括caC和fC提供了对表观遗传景观的更全面的理解.
- 最接近的邻基标识是影响修改的细胞因子测序精度的关键因素.
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