Wöhl T, Brecht M, Lottspeich F, Ammer H
Max-Planck-Institut für Biochemie, Martinsried, Germany.
Electrophoresis. 1995 May;16(5):739-41. doi: 10.1002/elps.11501601120.
Hybridization within agarose gels using oligonucleotide probes has been described in several publications; genomic DNA probes, however, have been used rarely and only with limited success. Here we present a simple and convenient procedure for in-gel hybridization using radiolabeled genomic DNA fragments. The protocol was improved by the use of formamide in the hybridization as well as in the washing step. This method was compared with the conventional Southern blotting technique and was shown to produce good results in restriction pattern analysis, as well as in chromosomal localization with the help of pulsed field gel electrophoresis.
琼脂糖凝胶内使用寡核苷酸探针的杂交技术已在多篇文献中有所描述;然而,基因组DNA探针的使用却很少,且成功率有限。在此,我们介绍一种使用放射性标记基因组DNA片段进行凝胶内杂交的简单便捷方法。通过在杂交及洗涤步骤中使用甲酰胺对该方案进行了改进。将此方法与传统的Southern印迹技术进行比较,结果表明,在限制性图谱分析以及借助脉冲场凝胶电泳进行染色体定位方面,该方法均能产生良好的结果。