Shiizaki Kazuhiro
Department of Applied Biosciences, Faculty of Life Sciences, Toyo University, Itakura, Gunma, Japan.
Methods Mol Biol. 2020;2119:213-226. doi: 10.1007/978-1-0716-0323-9_18.
In DNA adduct analysis, the P-postlabeling technique is a powerful tool due to its high detection sensitivity. It is performed by enzymatic digestion of DNA samples, enrichment of the adduct nucleotides, and then 5'-labeling with P. This method is particularly useful for detection of bulky adducts. An additional advantage is that only a small amount of DNA is required for detecting DNA adducts. This chapter describes the experimental procedure for separation and detection of DNA adducts by polyacrylamide gel electrophoresis, which is an attractive method for visually assessing differences in adduct formation between samples.
在DNA加合物分析中，P-后标记技术因其高检测灵敏度而成为一种强大的工具。它通过对DNA样品进行酶切、加合物核苷酸富集，然后用P进行5'-标记来完成。该方法对于检测大分子加合物特别有用。另一个优点是检测DNA加合物仅需少量DNA。本章描述了通过聚丙烯酰胺凝胶电泳分离和检测DNA加合物的实验步骤，这是一种直观评估样品间加合物形成差异的有吸引力的方法。
