Gadau Jürgen
School of Life Sciences, Arizona State University, Tempe, AZ 85287, USA.
Cold Spring Harb Protoc. 2009 Jul;2009(7):pdb.prot5245. doi: 10.1101/pdb.prot5245.
Many different DNA isolation methods have been employed successfully in ants. Parameters such as the size and developmental stage of the specimen (egg, larvae, or adult) and the subsequent use of the DNA will mostly determine which method should be used. Ant body sizes range from minute (1-2 mm in length) to large (30 mm), and the volume of the initial digestion should be adjusted accordingly. Whereas workers usually have low concentrations of storage proteins and fat, queens and larvae can contain considerable amounts of these substances that can interfere with the subsequent use of the isolated DNA. Ants also have many glands in the head and abdomen, and the contents of these glands can also interfere with the successful application of polymerase chain reaction (PCR) or restriction digests of the isolated DNA. This protocol presents two DNA isolation methods that have worked reliably for a wide range of ant species: a "quick and dirty" technique using Chelex isolation, and a more elaborate, classical phenol:chloroform procedure.
许多不同的DNA分离方法已在蚂蚁中成功应用。诸如标本的大小和发育阶段（卵、幼虫或成虫）以及DNA的后续用途等参数，大多会决定应使用哪种方法。蚂蚁的体型大小从微小（体长1-2毫米）到大型（30毫米）不等，初始消化的体积应相应调整。工蚁通常储存蛋白和脂肪的浓度较低，而蚁后和幼虫可能含有大量此类物质，这些物质会干扰分离出的DNA的后续使用。蚂蚁头部和腹部还有许多腺体，这些腺体的内容物也会干扰聚合酶链反应（PCR）的成功应用或分离出的DNA的限制性酶切。本方案介绍了两种已在多种蚂蚁物种中可靠应用的DNA分离方法：一种使用Chelex分离的“快速简便”技术，以及一种更精细的经典苯酚：氯仿法。
