RAPID AND QUANTITATIVE PREPARATION OF CYTOPLASMIC RNA FROM SMALL NUMBERS OF CELLS
RAPID AND QUANTITATIVE PREPARATION OF CYTOPLASMIC RNA FROM SMALL NUMBERS OF CELLS
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DOI：
10.1016/0003-2697(88)90164-9
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发表时间：
1988-08-15
影响因子：
2.9
通讯作者：
GOUGH, NM
中科院分区：
文献类型：
--
作者：
GOUGH, NM
描述了一种从少量细胞中制备胞质RNA的极其简单的方法。用去污剂NP-40裂解细胞，并通过十二烷基硫酸钠和尿素变性确保从核后细胞质裂解物中有效提取蛋白质。该方法适用于从许多细胞类型制备RNA。所有程序都已按比例缩小到在1.5 ml微量离心管中进行，因此可以从少量细胞制备RNA。该过程非常快速，RNA可用于北方凝胶分析，不到30分钟。由于涉及的步骤很少，RNA回收是定量的。
An extremely simple procedure for preparing cytoplasmic RNA from small numbers of cells is described. Cells are lysed with the detergent NP-40 and efficient extraction of protein from the postnuclear cytoplasmic lysate is ensured by denaturation with sodium dodecyl sulfate and urea. This procedure is suitable for preparing RNA from many cell types. All procedures have been scaled down to be performed in 1.5-ml microfuge tubes and thus RNA may be prepared from small numbers of cells. The procedure is extremely rapid and RNA is ready for Northern gel analysis is less than 30 min. Because so few steps are involved, RNA recovery is quantitative.