Plasmid DNA alkali cracking equipment and use method thereof

Abstract

The invention discloses plasmid DNA alkali cracking equipment. The equipment comprises a bacterial liquid pipeline; a buffer solution pipeline; a bacteria liquid suspension circulating device comprising circulating pipelines which are connected end to end to form a loop, and connected with the bacteria liquid pipeline and the buffer solution pipeline, wherein a pump and a valve arranged on the circulating pipeline; an alkali lysate pre-preparation pipeline; a lysis reactor; a neutralization reactor; an acid liquor pipeline; and a collecting device. The invention also discloses a method for extracting plasmid DNA by using the method. The method comprises the following steps: resuspending thalli by using a bacterial liquid suspension circulating device, pre-preparing an alkali lysate, breaking cells, inputting an acid solution, neutralizing a bacterial liquid and the like. The bacterial liquid can repeatedly circulated in the bacterial liquid suspension circulating device, the flow is improved, and the bacterial liquid resuspension effect is improved; the flow is adjusted, and thus the bacterial liquid can reach the target flow, and is convenient to mix with the alkali lysate.

Description

technical field [0001] The present invention relates to the production of biomolecules, and in particular to the field of polynucleotides such as plasmid DNA. Background technique [0002] In the prior art, alkaline lysis of plasmid DNA consists of three stages: the first stage is cell resuspension, which usually uses manual stirring or magnetic stirrer to resuspend cells in resuspension buffer; the second stage is cell lysis , lysis is usually performed by manual rotation or magnetic stirring to mix the resuspended cells with the lysis solution (consisting of diluted alkali (alkali) and detergent); the mixture is then kept at room temperature (20-25 degrees Celsius) or on ice A period of time, such as 5 minutes, to complete lysis; the third stage is neutralization and precipitation of host contaminants. Lysates from the second stage are usually mixed with cold neutralizing solution by gentle rotation or magnetic stirring to acidify the lysate, then placed on ice for 10-30 ...

AI Technical Summary

Problems solved by technology

Although the method and the equipment can be scaled up, the equipment cannot realize the continuous resuspension of bacteria, the pre-preparation of alkaline solution and the pre-starting of acid solution, so the solution (bacteria solution, acid, lye) cannot be realized according to the process. configuration, so that automatic continuous production and adaptation to different process parameters cannot be realized, so the production cycle cannot be shortened and the production cost can be increased

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