2琼脂糖凝胶电泳.pptVIP

* * Agarose Gel Electrophoresis 琼脂糖凝胶电泳 * * After the experiment is finished, students should know how to make an agarose gel. 掌握琼脂糖凝胶电泳的操作的方法。 一、实验目的 (Experimental Purpose) * * Gel electrophoresis * * 二、实验原理(Experimental Principle) Electrophoresis is a technique used to separate and sometimes purify proteins and nucleic acids, which differ in size, charge or conformation from each other. Electrophoresis has become the most widely used techniques in biochemistry and molecular biology. 电泳常用于分离和纯化那些分子大小、电荷性状或分子构象有所不同的生物大分子——尤其是蛋白质和核酸。正因为如此，电泳已成为生物化学和分子生物学中应用最为广泛的技术之一。 * * Agarose is a polysaccharide extracted from seaweed. Agarose gels have a large range of separation, but relatively low resolving power. By varying the concentration of agarose, fragments of DNA from about 200 to 50,000 bp can be separated using standard electrophoretic techniques. 琼脂糖是一种海藻多糖，琼脂糖胶分离范围很大，但其分辨率却相对较低。通过改变琼脂糖凝胶的浓度，应用标准的电泳技术可以分离200到50,000 bp 大小的 DNA 片断。 琼脂糖凝胶浓度越大，凝胶就越硬。较高浓度的琼脂糖胶有利于较小的DNA片断分离，而较低浓度的琼脂糖胶则可以分离较大的DNA片断。 The higher the agarose concentration, the stiffer the gel. Higher concentrations of agarose help in the separation of small DNAs, while low agarose concentrations allow resolution of larger DNAs. * * Agarose is typically used at concentrations of 0.5% to 2%. The distance DNA has migrated in the gel can be judged by visually monitoring migration of the tracking dyes. 琼脂糖胶浓度一般在0.5％到2％之间。 通过观察示踪染料的迁移距离可以判断DNA的迁移距离。溴酚蓝染料在琼脂糖凝胶的迁移速率最大。 当迁移足够距离后，就可以通过Gelview染色来观察DNA片断。 When adequate migration has occured, DNA fragments can be visualized by staining with Gelview. * * 琼脂糖凝胶浓度 0.3% 0.6% 0.9% 1.2% 1.5% 2% DNA片段分离范围 6-60 kb 1-20 kb 0.5-7kb 0.4-6kb 0.2-3kb 0.1-2kb * * Gelview is a fluorescent dye. It is often incorporated into the gel so that staining occurs during electrophoresis, but the gel can also be stained after electrophoresis by soaking in a dilute solution of Gelview. To visualize DNA or RNA, the gel is placed on a ul