Alkaline lysis system for preparing plasmid DNA and combined system

Abstract

The invention provides an alkaline lysis system for preparing plasmid DNA and a combined system. The alkaline lysis system for preparing plasmid DNA contains a bacterial suspension storage tank, an alkaline lysate storage tank and a hollow ultrafiltration column, wherein the hollow ultrafiltration column contains a first cavity and a second cavity which are divided by an ultrafiltration membrane;the alkaline lysate storage tank is communicated with the first cavity; and the bacterial suspension storage tank is communicated with the second cavity. The alkaline lysis system provided by the invention adopts the hollow ultrafiltration column to mix bacterial suspension and alkaline lysate evenly and avoid the problems that the pH value is too high locally and the shear stress is too large owning to the conventional mixing method such as stirring; and the invention provides the alkaline lysis system for preparing plasmid DNA, and the system has good stability, high efficiency and high repeatability. Meanwhile, the plasmid DNA prepared by the alkaline lysis system has higher supercoiled DNA content which is up to more than 90%.

Description

technical field [0001] The invention relates to the technical field of alkaline lysis, in particular to an alkaline lysis system and a combined system for preparing plasmid DNA. Background technique [0002] Gene therapy is in the research stage, and it may become a treatment for some major diseases such as tumors, atherosclerosis, osteoporosis, arthritis, etc., and the key to gene therapy is whether the carrier carrying the therapeutic gene can effectively enter the target area. cell. At present, the vectors used in gene therapy can be roughly divided into two categories, one is viral vectors and the other is non-viral vectors. At present, clinically, adenoviral vectors account for 24%, retroviral vectors account for 21.1%, and plasmid vectors account for 18%. Although the transfection rate of viral vectors is high, there are some safety hazards; while the gene expression efficiency of plasmid DNA in target cells is low and the duration is short, but it has high safety in...

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Problems solved by technology

[0004] The present invention aims to provide an alkali lysis system for preparing plasmid DNA, to solve the problem that the shear force brought by stirring in the alkali lysis reaction in the prior art cannot be controlled and the local pH is too high caused by uneven stirring, and further Provide a system suitable for industrial continuous production

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