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DOI:	10.1371/journal.pone.0148023
论文题名:	Critical Parameters for Efficient Sonication and Improved Chromatin Immunoprecipitation of High Molecular Weight Proteins
作者:	Nikolay A. Pchelintsev; Peter D. Adams; David M. Nelson
刊名:	PLOS ONE
ISSN:	1932-6203
出版年:	2016
发表日期:	2016-1-28
卷:	11, 期:1
语种:	英语
英文关键词:	Sonication ; Chromatin ; DNA purification ; Cross-linking ; Gel electrophoresis ; Ice core ; Polymerase chain reaction ; Lysis (medicine)
英文摘要:	Solubilization of cross-linked cells followed by chromatin shearing is essential for successful chromatin immunoprecipitation (ChIP). However, this task, typically accomplished by ultrasound treatment, may often become a pitfall of the process, due to inconsistent results obtained between different experiments under seemingly identical conditions. To address this issue we systematically studied ultrasound-mediated cell lysis and chromatin shearing, identified critical parameters of the process and formulated a generic strategy for rational optimization of ultrasound treatment. We also demonstrated that whereas ultrasound treatment required to shear chromatin to within a range of 100–400 bp typically degrades large proteins, a combination of brief sonication and benzonase digestion allows for the generation of similarly sized chromatin fragments while preserving the integrity of associated proteins. This approach should drastically improve ChIP efficiency for this class of proteins.
URL:	http://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0148023&type=printable
Citation statistics:
资源类型:	期刊论文
标识符:	http://119.78.100.158/handle/2HF3EXSE/23837
Appears in Collections:	过去全球变化的重建影响、适应和脆弱性科学计划与规划气候变化与战略全球变化的国际研究计划气候减缓与适应气候变化事实与影响

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journal.pone.0148023.PDF（2903KB）	期刊论文	作者接受稿	开放获取	CC BY-NC-SA	View Download

作者单位:

CR-UK Beatson Labs, Institute of Cancer Sciences, University of Glasgow, Glasgow, United Kingdom;CR-UK Beatson Labs, Institute of Cancer Sciences, University of Glasgow, Glasgow, United Kingdom;CR-UK Beatson Labs, Institute of Cancer Sciences, University of Glasgow, Glasgow, United Kingdom

Recommended Citation:

Nikolay A. Pchelintsev,Peter D. Adams,David M. Nelson. Critical Parameters for Efficient Sonication and Improved Chromatin Immunoprecipitation of High Molecular Weight Proteins[J]. PLOS ONE,2016-01-01,11(1)

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