Cockerill P N
Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia.
Anal Biochem. 1988 Feb 1;168(2):451-4. doi: 10.1016/0003-2697(88)90342-9.
Two methods which permit detection by autoradiography of small 32P-labeled DNA fragments resolved by agarose gel electrophoresis are described. Agarose gel electrophoresis poses problems for autoradiography as (i) the gels are normally too thick to allow autoradiography without being dried first, and (ii) fragments of DNA of 1000 bp or less in length are readily lost during drying. In this study DNA fragments as small as 121 bp have been retained in agarose gels upon drying. This has been achieved by either (i) first fixing the DNA with the cationic detergent cetyltrimethylammonium bromide, or (ii) drying the agarose gels onto Zeta-Probe charge-modified membranes.
本文描述了两种通过放射自显影检测经琼脂糖凝胶电泳分离的小的32P标记DNA片段的方法。琼脂糖凝胶电泳给放射自显影带来了一些问题，因为：（i）凝胶通常太厚，不先干燥就无法进行放射自显影；（ii）长度为1000bp或更短的DNA片段在干燥过程中很容易丢失。在本研究中，小至121bp的DNA片段在干燥后仍保留在琼脂糖凝胶中。这是通过以下两种方法实现的：（i）先用阳离子去污剂十六烷基三甲基溴化铵固定DNA；（ii）将琼脂糖凝胶干燥到Zeta-Probe电荷修饰膜上。
