Micard D, Sobrier M L, Couderc J L, Dastugue B
Anal Biochem. 1985 Jul;148(1):121-6. doi: 10.1016/0003-2697(85)90636-0.
A procedure for extracting RNA-free plasmid DNA from bacterial cells is described. The method is simple and rapid enough to obtain pure plasmid DNA in 8 to 10 h after plasmid amplification. The protocol uses the alkaline extraction procedure described by Birnboim and Doly (1979, Nucl. Acid Res. 7, 1513-1523). Plasmid DNA is then separated from high-molecular-weight RNA by ammonium acetate precipitation and from low-molecular-weight RNA contaminants by Ultrogel A2 column chromatography. The plasmid DNA obtained by this inexpensive technique is sufficiently pure to be used for restriction endonuclease analysis, 5'-end labeling, S1 mapping, DNA sequencing, and colony hydridization.
本文描述了一种从细菌细胞中提取无RNA质粒DNA的方法。该方法简单快速，在质粒扩增后8至10小时内即可获得纯质粒DNA。该方案采用了Birnboim和Doly（1979年，《核酸研究》7卷，1513 - 1523页）所述的碱提取法。然后通过醋酸铵沉淀从高分子量RNA中分离出质粒DNA，并通过Ultrogel A2柱色谱法从低分子量RNA污染物中分离出来。通过这种廉价技术获得的质粒DNA纯度足以用于限制性内切酶分析、5'端标记、S1图谱分析、DNA测序和菌落杂交。
