RAPID PURIFICATION OF PLASMID DNAS BY HYDROXYAPATITE CHROMATOGRAPHY
RAPID PURIFICATION OF PLASMID DNAS BY HYDROXYAPATITE CHROMATOGRAPHY
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DOI：
10.1111/j.1432-1033.1978.tb20966.x
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发表时间：
1978-01-01
期刊：
影响因子：
--
通讯作者：
LYONS, A
中科院分区：
文献类型：
--
作者：
COLMAN, A;BYERS, MJ;LYONS, A
A method is described for the rapid preparation of [Escherichia coli] plasmid DNA of MW up to 14 .times. 106. This method involves the chromatography, at room tempeature, of bacterial cleared lysates on hydroxyapatite in the presence of high concentrations of phosphate and urea. All detectable protein and RNA contamination of plasmid DNA is removed by this procedure and the conformation of the plasmid DNA is unaffected. Less than 0.5% chromosomal DNA is present in the purified preparation and even this can be removed if necessary by a simple extention of the procedure to include a heat-denaturation step. The method is extremely rapid and amenable to large-scale plasmid preparation; 5 mg ColE1 DNA were purified within 40 min. The yield of plasmid DNA is similar to that obtained with the conventional dye-centrifugation technique, but the purity is greater.
