一种使用毛细管电泳遗传分析仪进行DNA酶I足迹分析的推荐工作流程。

A recommended workflow for DNase I footprinting using a capillary electrophoresis genetic analyzer.

作者信息

Sivapragasam Smitha, Pande Anuja, Grove Anne

机构信息

Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA.

出版信息

Anal Biochem. 2015 Jul 15;481:1-3. doi: 10.1016/j.ab.2015.04.013. Epub 2015 Apr 20.

DOI:10.1016/j.ab.2015.04.013

PMID:25908559

Abstract

Fragment analysis was developed to determine the sizes of DNA fragments relative to size standards of known lengths using a capillary electrophoresis genetic analyzer. This approach has since been adapted for use in DNA footprinting. However, DNA footprinting requires accurate determination of both fragment length and intensity, imposing specific demands on the experimental design. Here we delineate essential considerations involved in optimizing the fragment analysis workflow for use in DNase I footprinting to ensure that changes in DNase I cleavage patterns may be reliably identified.

摘要

片段分析技术是为了利用毛细管电泳基因分析仪，相对于已知长度的大小标准来确定DNA片段的大小而开发的。从那以后，这种方法已被应用于DNA足迹分析。然而，DNA足迹分析需要精确测定片段长度和强度，这对实验设计提出了特定要求。在这里，我们阐述了优化用于DNase I足迹分析的片段分析工作流程所涉及的基本考虑因素，以确保能够可靠地识别DNase I切割模式的变化。