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Sanger Sequencing

Sanger Sequencing

DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...

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Updated: Jun 7, 2025

Targeted DNA Methylation Analysis by Next-generation Sequencing

Targeted DNA Methylation Analysis by Next-generation Sequencing

Published on: February 24, 2015

通过酶转化 (EM-seq) 进行全基因组甲基化测序:协议,数据处理和分析.

Nelly N Olova^1,2, Simon Andrews³

¹MRC Human Genetics Unit, MRC Institute of Genetics and Cancer, University of Edinburgh, Edinburgh, UK. nelly.olova@ed.ac.uk.

Methods in molecular biology (Clifton, N.J.)

|November 15, 2024

概括

酵素甲基化测序 (EM-seq) 为DNA甲基化分析提供了对全基因组双硫酸盐测序 (WGBS) 的无偏差替代方案. 这种新方法提供了精确的甲基化检测,减少了DNA输入和更少的文物.

关键词:

这是APOBECAPOBECAPOBECAPOBECAPOBECAPOBECAPOBECAPOBECAPOBECAPOBECAPOBECAPOBECAPOBEC 这就是BS-seqqq.比斯马克 (Bismark) 是一个国家.通过DNA甲基化.在EM-seqqq.酶性甲基组序列测序的方法下一个流程下一个流程这就是SeqMonk.在T4-BGT中使用.在TET2的基础上.这是WGBS的WGBS.

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Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

Published on: February 24, 2015

DNA Methylation: Bisulphite Modification and Analysis

DNA Methylation: Bisulphite Modification and Analysis

Published on: October 21, 2011

相关实验视频

Last Updated: Jun 7, 2025

Targeted DNA Methylation Analysis by Next-generation Sequencing

Targeted DNA Methylation Analysis by Next-generation Sequencing

Published on: February 24, 2015

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

Published on: February 24, 2015

DNA Methylation: Bisulphite Modification and Analysis

DNA Methylation: Bisulphite Modification and Analysis

Published on: October 21, 2011

科学领域:

表观遗传学在表观遗传学中,表观遗传学是指表观遗传学.
基因组学就是基因组学.
分子生物学分子生物学

背景情况:

15年来,全基因组双硫酸盐测序 (WGBS) 是DNA甲基化分析的标准.
WGBS容易出现技术偏差,导致过高估计的甲基化值和氨酸丰富地区的文物.
双硫酸盐的转化是对DNA的破坏性过程.

研究的目的:

介绍和评估酶甲基化测序 (EM-seq) 作为WGBS的优质替代方案.
强调EM-seq用于DNA甲基化分析的优点.
证明EM-seq.的无偏差和无退化性质.

主要方法:

在EM-seq中采用了使用TET2,T4-BGT和APOBEC的两步酶转化过程.
细胞因子通过氧化和葡萄糖化得到保护.
未经修改的细胞因子被去胺化为 uracil.

主要成果:

与WGBS不同,EM-seq是没有降解和无偏差的.
该技术需要较低的DNA输入,并产生较高的库输出,读取时间较长.
EM-seq在更多的CPG中展示了统一的基因组覆盖,减少重复和准确的甲基化检测.
使用EM-seq.没有观察到特定序列的工件.

结论:

EM-seq代表了DNA甲基化分析的重大进步,取代了破坏性WGBS方法.
该技术为基础分辨率甲基化研究提供了提高的准确性,效率和可靠性.
EM-seq是一种强大的,无偏见的方法,适合各种基因组应用,需要精确的甲基化概况.