Gupta R C
Preventive Medicine and Environmental Health, University of Kentucky, Lexington 40506.
IARC Sci Publ. 1993(124):11-23.
The 32P-postlabelling assay has emerged as a major tool for detecting DNA adducts induced by structurally diverse carcinogens, particularly bulky aromatics. The assay comprises enzymatic degradation of DNA to 3'-mononucleotides, enrichment of adducts, 5'-32P-labelling, adduct separation by TLC, and detection and quantitation of adducts. This report describes (1) an improved solvent extraction procedure for isolating DNA from tissues and (2) an up-dated version of the assay that we consider optimal for bulky adducts. The use of a non-urea solvent mixture (isopropanol: 4 M ammonium hydroxide) has been found to improve adduct separation and signal-to-noise ratios issues and (2) an up-dated version of the assay that we consider optimal for bulky adducts. The use of a non-urea solvent mixture (isopropanol: 4 M ammonium hydroxide) has been found to improve adduct separation and signal-to-noise ratios.
32P后标记分析法已成为检测由结构多样的致癌物（尤其是大分子芳烃）诱导产生的DNA加合物的主要工具。该分析方法包括将DNA酶解为3'-单核苷酸、加合物富集、5'-32P标记、通过薄层色谱法分离加合物以及加合物的检测和定量。本报告描述了（1）一种从组织中分离DNA的改进溶剂萃取程序，以及（2）我们认为对大分子加合物最为适用的该分析方法的更新版本。已发现使用非尿素溶剂混合物（异丙醇：4M氢氧化铵）可改善加合物分离及信噪比。
