Plasmid DNA alkali cracking equipment and use method thereof

A technology of alkaline lysis and plasmids, which is applied in the direction of DNA preparation, biochemical equipment and methods, recombinant DNA technology, etc., can solve the problems of increasing production costs, not being able to shorten the production cycle, and not being able to achieve continuous bacterial resuspension, so as to shorten production cycle, increase production cost, and facilitate the effect of neutralization reaction

Active Publication Date: 2020-10-02
LISUI TECH SUZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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

Method used

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  • Plasmid DNA alkali cracking equipment and use method thereof
  • Plasmid DNA alkali cracking equipment and use method thereof

Examples

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Embodiment 1

[0031] figure 1 Schematically shows a plasmid DNA alkaline lysis device according to one embodiment of the present invention.

[0032] Such as figure 1 , the plasmid DNA alkaline lysing equipment includes a bacterial liquid pipeline 1, the pipe diameter is preferably 3.2mm, and the bacterial liquid pipeline is provided with a pinch valve and a peristaltic pump 14; the buffer pipeline 2, with a pipe diameter of preferably 6.4mm, is provided with a pinch valve And peristaltic pump 14; Buffer pipeline 2 connects bacterial solution pipeline 1 to mix bacterial solution through T connector or Y connector pipeline, also can directly connect bacterial solution suspension circulating device 6 and mix bacterial solution in circulation pipeline 61, connect The pipeline is provided with a pinch valve; in this embodiment, the mixed bacteria solution pipeline 5 formed after the buffer pipeline 2 is connected to the bacteria solution pipeline 1 is connected to the inlet of the circulation p...

Embodiment 2

[0042] figure 2 A method for extracting plasmid DNA according to one embodiment of the present invention is schematically shown.

[0043] Such as figure 2 As shown, the method includes the following steps:

[0044] Start the bacteria solution pipeline 1 and the buffer solution pipeline 2, transport the bacteria and buffer solution to the bacteria solution suspension circulation device 6 at a volume ratio of 1:10 through the peristaltic pump 14 and the flow sensor, and close the bacteria solution pipeline 1 after reaching the preset volume and buffer line 2.

[0045] Start the bacterium liquid suspension circulation device 6, and the bacterium mixed liquid circulates in the loop that circulation pipe 61, stirring container 62 constitute; up to the target flow.

[0046] Alkaline lysate is pre-configured: start buffer pipeline 2, lye delivery pipeline 31 and surfactant delivery pipeline 32, alkali lysis solution pipeline 33 communicates with waste discharge pipeline 9, clos...

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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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12N15/10
CPCC12N15/1003
Inventor 周胜徐慧刘波高延林贾伟
Owner LISUI TECH SUZHOU
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