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DNA 5-甲基胞嘧啶特异性扩增与测序。

DNA 5-Methylcytosine-Specific Amplification and Sequencing.

机构信息

Department of Chemistry, Department of Biochemistry and Molecular Biology, and Institute for Biophysical Dynamics, The University of Chicago, Chicago, Illinois 60637, United States.

Howard Hughes Medical Institute, The University of Chicago, Chicago, Illinois 60637, United States.

出版信息

J Am Chem Soc. 2020 Mar 11;142(10):4539-4543. doi: 10.1021/jacs.9b12707. Epub 2020 Feb 25.

DOI:10.1021/jacs.9b12707
PMID:32077696
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7216556/
Abstract

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 5mC-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.

摘要

DNA 5-甲基胞嘧啶（5mC）特异性作图受到常规亚硫酸氢盐测序方法中严重的 DNA 降解和 5-羟甲基胞嘧啶（5hmC）存在的阻碍。在这里，我们提出了一种 5mC 特异性全基因组扩增方法（5mC-WGA），通过该方法，我们实现了在有限的输入下（低至 10pg）进行 DNA 扩增时保留 5mC，同时受到 5hmC 信号的干扰有限，从而提供了具有高重复性和准确性的 DNA 5mC 甲基组。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be4f/7216556/75e5f893169a/nihms-1580469-f0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be4f/7216556/895b519c8636/nihms-1580469-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be4f/7216556/68624ffe33d2/nihms-1580469-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be4f/7216556/ec80b300e865/nihms-1580469-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be4f/7216556/75e5f893169a/nihms-1580469-f0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be4f/7216556/895b519c8636/nihms-1580469-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be4f/7216556/68624ffe33d2/nihms-1580469-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be4f/7216556/ec80b300e865/nihms-1580469-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be4f/7216556/75e5f893169a/nihms-1580469-f0004.jpg

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