Werner D, Chemla Y, Herzberg M
Anal Biochem. 1984 Sep;141(2):329-36. doi: 10.1016/0003-2697(84)90050-2.
Poly(A)+ RNA was isolated from in vitro short-term-labeled total cytoplasmic RNA of Ehrlich ascites tumor cells by oligo(dT) cellulose chromatography. This poly(A)+ RNA fraction was compared with a poly(A)+ RNA fraction isolated by a new procedure which involves specific binding of poly(A)+ RNA to messenger affinity paper (mAP) and its release in hot (70 degrees C) water. In typical experiments 10-11 micrograms (2.3%) of poly(A)+ RNA can be retained from 500 micrograms of total cytoplasmic RNA per cm2 of mAP in a quick one-step procedure. The poly(A)+ RNA preparations isolated by the two methods proved to be almost identical with respect to their fraction in total cytoplasmic RNA, specific radioactivities, sucrose gradient profiles, and translation assays. Since the isolation of poly(A)+ RNA by mAP is much less time consuming than that by oligo(dT) column chromatography and since the poly(A)+ RNA can be recovered from mAP in small volumes, which avoids further loss during precipitations, it can be advantageously used for preparative isolation of poly(A)+ RNA.
通过寡聚（dT）纤维素柱层析从艾氏腹水瘤细胞体外短期标记的总细胞质RNA中分离出聚腺苷酸（Poly(A)）+RNA。将该Poly(A)+RNA组分与通过一种新方法分离的Poly(A)+RNA组分进行比较，该新方法涉及Poly(A)+RNA与信使亲和纸（mAP）的特异性结合及其在热水（70℃）中的释放。在典型实验中，每平方厘米mAP可在快速一步法中从500微克总细胞质RNA中保留10 - 11微克（2.3%）的Poly(A)+RNA。通过两种方法分离的Poly(A)+RNA制剂在总细胞质RNA中的比例、比放射性、蔗糖梯度图谱和翻译分析方面几乎相同。由于通过mAP分离Poly(A)+RNA比通过寡聚（dT）柱层析耗时少得多，并且由于Poly(A)+RNA可以从小体积的mAP中回收，避免了沉淀过程中的进一步损失，因此它可有利地用于Poly(A)+RNA的制备性分离。
