The preparation of poly(A)+mRNA from the hagfish slime gland.
The preparation of poly(A)+mRNA from the hagfish slime gland.
复制标题
从盲鳗粘液腺中制备 Poly(A) mRNA。
DOI：
10.1080/00327488808062513
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Carlson,DM
中科院分区：
文献类型：
--
作者：
Gadbois,DM;Salo,WL;Ann,DK;Downing,SW;Carlson,DM
从太平洋盲鳗的粘液腺中分离可翻译的poly（A）±mRNA是不可能的，因为当腺体的内容物水合时形成粘性粘液，所以通常使用的方法是不可能的。本文报道了一个程序，以克服这个问题。简言之，将组织在液氮中研成粉末，与月桂酰肌氨酸钠和蛋白酶K混合并冻干。然后将冻干粉末与0.3 mm直径的玻璃珠混合，充分研磨并用缓冲液润湿，并在37°C下消化。通过CsCl垫进行超离心，从消化物中回收RNA。RNA的进一步纯化是通过常规方法进行的，并稍作修改。
The isolation of translatable poly(A)±mRNA from the slime glands of the Pacific hagfish,Eptatretus stouti, is not possible by the commonly used procedures because of the viscous slime that is formed when the contents of the glands are hydrated. This paper reports on a procedure developed to overcome this problem. Briefly, the tissue was powdered in liquid nitrogen, mixed with sodium lauroylsarcosine and proteinase K and lyophilized. The lyophilized powder was then mixed with 0.3 mm diameter glass beads, thoroughly ground and wetted with buffer and digested at 37°C. The RNA from the digest was recovered by ultracentrifugation through a CsCl cushion. Further purification of the RNA was accomplished by the usual methods with slight modifications.