Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

Dimethyl sulfate (DMS) footprinting assay

TB Tao Bing
WZ Wei Zheng
XZ Xin Zhang
LS Luyao Shen
XL Xiangjun Liu
FW Fuyi Wang
JC Jie Cui
ZC Zehui Cao
DS Dihua Shangguan
request Request a Protocol
ask Ask a question
Favorite

Dimethyl sulfate footprinting experiments were performed according to the protocol developed by Tan et al.58 10 picomoles of FAM-labeled CBA-0 or CBA-1 in 200 µL volume were mixed with 4 µL of 10% (v/v) dimethyl sulfate (DMS) in ethanol and incubated for 2 min at room temperature. The reaction was stopped by addition of 200 µL of  stop buffer (0.6 M NaOAc, 0.1 M β-mercaptoethanol, 20 µg sperm DNA). After phenol/chloroform extraction and ethanol precipitation, the DNA was dissolved in 50 µL of  water. Thereafter, 50 µL of  20% (v/v) piperidine in water was added and the samples were heated at 90 °C for 30 min, followed by phenol/chloroform extraction and ethanol precipitation. The precipitated DNA was dissolved in 50% (v/v) deionized formamide in water, denatured at 95 °C for 5 min and resolved on a denaturing 12% polyacrylamide gel.

Copyright and License information: The Author(s) ©2017
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A