Isolation procedure for bacterial DNA based on gel permeation chromatography on a Sepharose column

✍ Scribed by S. Zadražil; J. Šatava; Zora Šormová

• Book ID: 104146837
• Publisher: Elsevier Science
• Year: 1974
• Tongue: English
• Weight: 597 KB
• Volume: 91
• Category: Article
• ISSN: 1873-3778
• DOI: 10.1016/s0021-9673(01)97924-5

✦ Synopsis

Gel permeation chromatography of bacterial lysates, no matter what type of lysis is used, on a Sepharose 4B column equilibrated with buffered 2 M NaCl was demonstrated to be generally useful as a simple method for the quantitative and reproducible isolation of high-molecular-weight DNA (average molecular weight 2.2 x 10' daltons). Conventional analyses of this DNA and its re-chromatography on a MAK column and electrophoretic fractionations in polyacrylamide and agarose gels show less heterogeneity and a lower polysaccharide content in comparison with DNA preparations isolated by other complex processes, no RNA contamination and 2-3 % of residual proteins. The method is also recommended for bacteria affected by different factors, where classical methods for the quantitative isolation of DNA fail.