Ferrero E A, Fitch M M, Moore P H, Wu K K
Hawaiian Sugar Planters' Association Experimental Station, Aiea, USA.
Biotechniques. 1995 Feb;18(2):266-73.
Nonradioactive Southern blotting using digoxigenin (DIG) has become routinely applied to the analysis of single-copy genes for genetic mapping because it is fast and safe. Previous studies indicate that DIG-labeled probes are suitable for single-gene detection in less complex genomes, but their efficient application to mapping a large octoploid genome has not been discussed. We developed a stream-lined procedure for nonradioactive restriction fragment length polymorphism mapping and DNA fingerprinting of sugarcane that combines DIG-11-dUTP and anion-exchange chromatography. In this report, we show that anion-exchange chromatography provides a reliable and simple technique for the resin-purification of large numbers of DIG-labeled DNA fragments 0.3-3.0 kb in size, and it is essential in minimizing contaminants and nonspecific signal.
使用地高辛（DIG）的非放射性Southern印迹法已常规应用于单拷贝基因的遗传图谱分析，因为它快速且安全。先前的研究表明，DIG标记的探针适用于较简单基因组中的单基因检测，但尚未讨论其在大型八倍体基因组图谱绘制中的有效应用。我们开发了一种简化的程序，用于甘蔗的非放射性限制性片段长度多态性图谱绘制和DNA指纹分析，该程序结合了DIG-11-dUTP和阴离子交换色谱法。在本报告中，我们表明阴离子交换色谱法为大量大小在0.3 - 3.0 kb的DIG标记DNA片段的树脂纯化提供了一种可靠且简单的技术，并且对于最大限度地减少污染物和非特异性信号至关重要。
