Sparks R B, Elder J H
Anal Biochem. 1983 Dec;135(2):345-8. doi: 10.1016/0003-2697(83)90694-2.
A simple and efficient procedure for the rapid isolation of plasmid DNA free of chromosomal DNA and with only minor contamination with RNA is described. The protocol is a modification of the boiling method described by Holmes and Quigley [(1981) Anal. Biochem. 114, 193-197.] and utilizes C18 reverse-phase silica beads for final concentration and purification of plasmid DNA. The entire procedure can be carried out in 1 day and does not require the use of phenol or cesium chloride gradients, which require considerable labor and may sometimes cause nicking and lower recoveries of supercoiled DNA. The plasmid DNA obtained by this method retains biological activity, is supercoiled, and is suitable for restriction and DNA sequence analysis.
本文描述了一种简单有效的方法，可快速分离不含染色体DNA且仅受少量RNA污染的质粒DNA。该方案是对Holmes和Quigley[(1981)Anal.Biochem.114,193 - 197.]所描述的煮沸法的改进，利用C18反相硅胶珠对质粒DNA进行最终浓缩和纯化。整个过程可在1天内完成，无需使用苯酚或氯化铯梯度，而使用这些方法需要大量人力，有时还可能导致超螺旋DNA切口和回收率降低。通过该方法获得的质粒DNA保留生物活性，呈超螺旋结构，适用于限制性酶切和DNA序列分析。
