a one-step miniprep for the isolation of plasmid dna and lambda phage particles的一步miniprep隔离的质粒dna和λ噬菌体粒子.pdfVIP

A One-Step Miniprep for the Isolation of Plasmid DNA and Lambda Phage Particles 1 1 2 3 1 George Lezin , Yasuhiro Kosaka , H. Joseph Yost , Michael R. Kuehn *, Luca Brunelli * 1 Division of Neonatology, Department of Pediatrics, The University of Utah School of Medicine, Salt Lake City, Utah, United States of America, 2 Department of Neurobiology and Anatomy, The University of Utah School of Medicine, Salt Lake City, Utah, United States of America, 3 Laboratory of Protein Dynamics and Signaling, National Cancer Institute, National Institutes of Health, NCI-Frederick, Frederick, Maryland, United States of America Abstract Plasmid DNA minipreps are fundamental techniques in molecular biology. Current plasmid DNA minipreps use alkali and the anionic detergent SDS in a three-solution format. In addition, alkali minipreps usually require additional column-based purification steps and cannot isolate other extra-chromosomal elements, such as bacteriophages. Non-ionic detergents (NIDs) have been used occasionally as components of multiple-solution plasmid DNA minipreps, but a one-step approach has not been developed. Here, we have established a one-tube, one-solution NID plasmid DNA miniprep, and we show that this approach also isolates bacteriophage lambda particles. NID minipreps are more time-efficient than alkali minipreps, and NID plasmid DNA performs better than alkali DNA in many downstream applications. In fact, NID crude lysate DNA is sufficiently pure to be used in digestion and sequencing reactions. Microscopic analysis showed that the NID procedure fragments E.coli cells into small protoplast-like components, which may, at least in part, explain the effectiveness of this approach. This work demonstrates tha