Reineke A, Karlovsky P, Zebitz C P
Institute of Phytomedicine, University of Hohenheim, Stuttgart, Germany.
Insect Mol Biol. 1998 Feb;7(1):95-9. doi: 10.1046/j.1365-2583.1998.71048.x.
Analysis of amplified fragment length polymorphism (AFLP) has the potential to become a powerful new DNA fingerprinting technique for studying genetic relationships and genetic diversity in arthropods. Since DNA of high quality is a crucial prerequisite for AFLP analysis we evaluated the applicability of six protocols (one fast and four complex methods with phenol-chloroform treatments as well as one CTAB-based method) for extracting DNA from insect material and three additional DNA purification steps. The most rapid DNA isolation method did not produce DNA suitable for AFLP analysis. Among four complex methods tested, two protocols resulted in comparatively low yields of DNA that was therefore not used as template for AFLP analysis. The other two complex methods with phenol treatments and a CTAB-based DNA extraction protocol provided DNA suitable for AFLP assay. An additional purification of the DNA using spermine precipitation revealed a few extra bands in an AFLP gel that were masked in unpurified DNA. Therefore spermine precipitation is recommended for AFLP templates.
扩增片段长度多态性（AFLP）分析有潜力成为一种强大的新型DNA指纹技术，用于研究节肢动物的遗传关系和遗传多样性。由于高质量的DNA是AFLP分析的关键前提条件，我们评估了六种方案（一种快速方法、四种经过酚 - 氯仿处理的复杂方法以及一种基于CTAB的方法）从昆虫材料中提取DNA的适用性，以及另外三个DNA纯化步骤。最快的DNA分离方法所产生的DNA不适用于AFLP分析。在测试的四种复杂方法中，有两种方案得到的DNA产量相对较低，因此未用作AFLP分析的模板。另外两种经过酚处理的复杂方法和一种基于CTAB的DNA提取方案提供了适用于AFLP检测的DNA。使用精胺沉淀对DNA进行额外纯化后，在AFLP凝胶中显示出一些在未纯化DNA中被掩盖的额外条带。因此，推荐将精胺沉淀用于AFLP模板。
