Karve M S, Kale N R
Prep Biochem. 1977;7(5):403-14. doi: 10.1080/00327487708061657.
A simple procedure for the preparation of agarose suitable for electrophoresis is developed in which anionic polysaccharides are removed by extracting the agar gel-granules with phosphate buffer (0.03 M, pH 6.8) containing urea (4 M), followed by electrophoresis in the same buffer system. Further, alkali treatment in the presence of sodium borohydride, eliminates electroendosmosis, giving essentially a neutral agarose, as judged by the electrophoretic behaviour of basic substances like crystal violet and cytochrome C. The purified agarose with yields 60-65%, has a sulphur content less than 0.1%, and forms rigid, transparent gels.
开发了一种制备适用于电泳的琼脂糖的简单方法,其中通过用含有尿素(4M)的磷酸盐缓冲液(0.03M,pH6.8)提取琼脂凝胶颗粒来去除阴离子多糖,然后在相同的缓冲系统中进行电泳。此外,在硼氢化钠存在下进行碱处理可消除电渗,根据结晶紫和细胞色素C等碱性物质的电泳行为判断,得到基本上呈中性的琼脂糖。纯化后的琼脂糖产率为60 - 65%,硫含量低于0.1%,并形成坚硬、透明的凝胶。